Bridged Macrocyclic Module Compositions

ABSTRACT

This invention is related to the fields of organic chemistry and nanotechnology. In particular, it relates to materials and methods for the preparation of organic synthons and bridged macrocyclic module components. The bridge macrocyclic module compounds may be used to prepare macrocyclic module compositions such as nanofilms, which may be useful for filtration.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §119(e) of U.S.Provisional Application No. 60/492,808 filed on Aug. 6, 2003, thedisclosure of which is hereby incorporated by reference in its entirety.

TECHNICAL FIELD

This invention is related to the fields of organic chemistry andnanotechnology. In particular, it relates to materials and methods forthe preparation of organic synthons and bridged macrocyclic modulecompositions. The bridged macrocyclic module compositions may be used toprepare macromolecular compositions such as nanofilms, which may beuseful for filtration.

BACKGROUND OF THE INVENTION

One area of nanotechnology is to develop chemical building blocks fromwhich hierarchical macromolecules of predicted properties can beassembled. An approach to making chemical building blocks ornanostructures begins at the atomic and molecular level by designing andsynthesizing starting materials with highly tailored properties. Precisecontrol at the atomic level is the foundation for development ofrationally tailored synthesis-structure-property relationships which canprovide materials of unique structure and predictable properties. Thisapproach to nanotechnology is inspired by nature where, for example,from twenty common amino acids found in natural proteins, more than 10⁵stable and unique proteins are made.

Nanotechnology has also been described by K. Eric Drexler in Engines ofCreation as “the knowledge and means for designing, fabricating andemploying molecular scale devices by the manipulation and placement ofindividual atoms and molecules with precision on the atomic scale.” Aquest of nanotechnology is to prepare molecular architectures capable ofperforming on a nanometer scale functions normally observed forlarge-scale constructs. For example, rotaxanes and polyrotaxanes aremolecules that are interlocked, but not chemically bound to one another,which act like nano-machines. In other examples, carbon nanotubes andsimilar constructs have been created which may function as molecularscaffold units, or as transport channels, storage units, orencapsulators for various atoms and molecules. The use of biologicalprocesses is also being studied as an approach to the assembly ofnon-biological nano-devices.

A hurdle in developing “building block” nanotechnology is creating theability to program the final output of a chemical reaction between thereactants from which the building blocks are formed. In other words, itis desirable to control the geometry of the building block reactants inorder to predetermine which product will be built from the reactants.The product will be the thermodynamically favored product in most cases,however, the programming of geometrical constraints into the reactantsoverrides the random statistics of bulk phase macroscopic interactionsand effectively limits the reaction at the atomic level.

Entropically driven self assembly processes may be used to producehierarchical products which are typically well-organized aggregates.Although such processes may be robust in tolerating a range ofconditions, they are generally very limited in synthetic accessibilityand produce a narrow range of products.

Most conventional methods of preparation of organic compounds involvestepwise attachment of species to form a product. Step-by-step synthesiscan be an arduous route to prepare molecules, even some relativelysimple molecules. Random statistics plays a role in every step and canmake it impossible to achieve multistep products. For example, acollection of cyclic synthons may be coupled in stepwise synthesis toyield macrocyclic modules, however, the yield in many cases isrelatively low, or may even be prohibitive.

One application that will benefit from nanotechnology is filtrationusing membranes. Conventional membranes used in a variety of separationprocesses can be made selectively permeable to various molecularspecies. The permeation properties of conventional membranes generallydepend on the pathways of transport of species through the membranestructure. While the diffusion pathway in conventional selectivelypermeable materials can be made tortuous in order to control permeation,porosity is not well defined or controlled by conventional methods. Theability to fabricate regular or unique pore structures of membranes is along-standing goal of separation technology.

Thus, what is needed is an approach to making chemical entities in theform of bridged macrocyclic module compositions from which to createnanostructures with desirable properties.

All documents referenced herein, including applications for patent,patent references, publications, articles, books, and treatises, arespecifically incorporated by reference herein in their entirety.

BRIEF SUMMARY OF THE INVENTION

A bridged macrocyclic module compound of the formula:

wherein the compound further comprises a bridge moiety A having two ormore termini, wherein at least two of said two or more termini arecoupled to the compound; wherein each Q is a synthon independentlyselected from the group consisting of benzene, cyclohexadiene,cyclopentadiene, naphthalene, anthracene, phenylene, phenanthracene,pyrene, triphenylene, phenanthrene, pyridine, pyrimidine, pyridazine,biphenyl, bipyridyl, cyclohexane, cyclohexene, decalin, piperidine,pyrrolidine, morpholine, piperazine, pyrazolidine, quinuclidine,tetrahydropyran, dioxane, tetrahydrothiophene, tetrahydrofuran, pyrrole,cyclopentane, cyclopentene, triptycene, adamantane,bicyclo[2.2.1]heptane, bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane,bicyclo[2.2.2]octene, bicyclo[3.3.0]octane, bicyclo[3.3.0]octene,bicyclo[3.3.1]nonane, bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane,bicyclo[3.2.2]nonene, bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, spiro[4.4]nonane, —OCH₂CH₂—,—(CH₂)_(n)C≡C(CH₂)_(n)—, —(CH₂)_(n)CH═CH(CH₂)_(n)—,

—(CH₂)_(n)—, —C(O)O(CH₂)_(n)—, —(CH₂)_(n)C(O)NR—; —S_(m)—,—(CH₂)_(n)SiMe₂(CH₂)_(n)—, —(CH₂)_(n)NR(CH₂)_(n)—, and—(CH₂)_(n)CH(OH)—; wherein each synthon Q may optionally be substitutedwith one or more functional groups for coupling the synthon to at leasta second bridged macrocyclic module or to a substrate; wherein eachsynthon may optionally be substituted with one or more lipophilic and/orhydrophilic groups; wherein each L is a linkage moiety independentlyselected from the group consisting of a direct bond, —NRC(O)—, —OC(O)—,—O—, —S—S—, —S—, —NR—, —(CRR)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—,—C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—,—NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—NH(CH₂)_(h)CH═N—, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—,—CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

wherein the linkage is independently configured in either of twopossible configurations, forward and reverse, with respect to thesynthons it couples together; wherein the bridge moiety A is selectedfrom the group consisting of:

—O—(CH₂)_(m)—O—; —{NH—CHR—(CO)}_(m)—O—; —O—(CF₂)_(m)—O—; —(S)_(m)—;—O(CH₂CH₂O)_(m)—; —(OCH(CH₃)CH₂)_(m)O—;

wherein the at least two termini of the bridge moiety may be conjugatedto the compound through a linkage moiety L, wherein L is as definedabove; wherein R is independently selected from the group consisting ofhydrogen and alkyl; wherein Ph is phenyl; wherein X is selected from thegroup consisting of F, Cl, Br, and I; wherein X′ is H or a functionalgroup for linking to at least a second bridged macrocyclic moiety or asubstrate; wherein each R² is independently selected from a bond forlinking to a synthon or a functional group selected from the groupconsisting of hydrogen, an activated acid, —OH, —C(O)OH, —C(O)H,—C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa, —SH,—C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂,—CH═CHR, —CH═CRR, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

—P(O)(OH)(OX), or —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein n′ is from 4 to 50;wherein n is 1-22; wherein m is 2-14; wherein p is 1-6; wherein h is1-4; wherein r is 1-50; and wherein s is 1-4.

In one embodiment, the two or more termini of said bridge moiety arecoupled to synthons. In another embodiment, the two or more termini ofsaid bridge moiety are coupled to L moieties, with the proviso that saidL moieties to which the termini are coupled are not direct bonds.

In one embodiment, n is from 4 to 24. In another embodiment, n is from 6to 16. In another embodiment, n is from 6 to 12. In other embodiments, nmay be, for example, 4, 6, 8, 10, 12, 14, or 16.

In another embodiment, the bridged macrocyclic module compound has theformula:

wherein Q¹, Q² and Q³ are as defined for Q above, L is as defined above,and A is a bridge moiety as defined above. In another embodiment, thebridged macrocyclic module compound has the formula:

wherein Q¹, Q² and Q³ are as defined for Q above, L is as defined above,and A is a bridge moiety as defined above. In another embodiment, thebridged macrocyclic module compound has the formula:

wherein Q¹, Q² and Q³ are as defined for Q above, L is as defined above,and A is a bridge moiety as defined above.

In another embodiment, each Q¹ is the same synthon. In anotherembodiment, each Q² is the same synthon. In another embodiment, each Q³is the same synthon.

In another embodiment, Q¹, Q², and Q³ are synthons independentlyselected from the group consisting of

wherein each X′ is independently H or a functional group for couplingthe synthon to at least a second bridged macrocyclic module or to asubstrate, wherein each J is an independently selected functional groupfor coupling the synthon to an adjacent synthon within said bridgedmacrocyclic module, and wherein each X₁ is an independently selectedfunctional group which may couple the synthon to the bridge moiety. Inanother embodiment, each Q¹, Q², and Q³ is independently selected fromthe group consisting of

wherein each X′ is independently H or a functional group for couplingthe synthon to at least a second bridged macrocyclic module or to asubstrate, wherein each J is an independently selected functional groupfor coupling the synthon to an adjacent synthon within said bridgedmacrocyclic module, and wherein each X₁ is an independently selectedfunctional group which may couple the synthon to the bridge moiety. In apreferred embodiment, each Q¹, Q², and Q³ is independently selected fromthe group consisting of

In another embodiment, each linker moiety L between the synthons is thesame. In another embodiment, each linker moiety L between the bridgemoiety and the synthons is the same.

In another embodiment, the synthons are cyclic synthons. In anotherembodiment, the synthons are acyclic synthons. In another embodiment,the synthons alternate between cyclic and acyclic synthons. In oneembodiment, the synthons are independently selected from the groupconsisting of benzene, cyclohexadiene, cyclopentadiene, naphthalene,anthracene, phenylene, phenanthracene, pyrene, triphenylene,phenanthrene, pyridine, pyrimidine, pyridazine, biphenyl, bipyridyl,cyclohexane, cyclohexene, decalin, piperidine, pyrrolidine, morpholine,piperazine, pyrazolidine, quinuclidine, tetrahydropyran, dioxane,tetrahydrothiophene, tetrahydrofuran, pyrrole, cyclopentane,cyclopentene, triptycene, adamantane, bicyclo[2.2.1]heptane,bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane, bicyclo[2.2.2]octene,bicyclo[3.3.0]octane, bicyclo[3.3.0]octene, bicyclo[3.3.1]nonane,bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane, bicyclo[3.2.2]nonene,bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, spiro[4.4]nonane. In another embodiment,the synthons are independently selected from the group consisting of—OCH₂CH₂—, —(CH₂)_(n)C≡C(CH₂)_(n)—, —(CH₂)_(n)CH═CH(CH₂)_(n)—,

—(CH₂)_(n)—, —C(O)O(CH₂)_(n)—, —(CH₂)_(n)C(O)NR—; —S_(m)—,—(CH₂)_(n)SiMe₂(CH₂)_(n)—, —(CH₂)_(n)NR(CH₂)_(n)—, and—(CH₂)_(n)CH(OH)—. In a preferred embodiment, the synthons areindependently selected from benzene, piperidine, and cyclohexane. Inanother preferred embodiment, the synthons are independently selectedfrom benzene and cyclohexane.

In one embodiment, each L is a bond. In another embodiment, L is alinkage independently selected from the group consisting of —NRC(O)—,—OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR)_(p)—, —CH₂NH—, —CH═N—, —C(O)S—,—C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—,—NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—N═CH(CH₂)_(h)CH═N—, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—,—CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

wherein p is 1-6; wherein h is 1-4; wherein Ph is phenyl; wherein each Ris independently selected from the group of hydrogen and alkyl; whereinthe linkage is independently configured in either of two possibleconfigurations, forward and reverse, with respect to the synthons itcouples together. In a preferred embodiment, each L is independentlyselected from the group consisting of —NH—C(O)—, —NH₂—, and —NH—CH₂—. Inone embodiment, L is not —CH₂—. In another embodiment, L is not—CH₂NHCO—.

In one embodiment, the bridge moiety further comprises a surfaceattachment group. In another embodiment, the bridge moiety furthercomprises a lipophilic group. In another embodiment, the bridge moietycomprises a functional group for coupling the compound to at least asecond bridged macrocyclic module compound. In another embodiment, thebridge moiety comprises a polymerization center. In one embodiment, thebridge moiety is not a polyethylene glycol moiety. In anotherembodiment, the bridge moiety does not comprise ethylene glycolmoieties. In one embodiment, X′ is H. In another embodiment, X′ is afunctional group.

In a preferred embodiment, the bridge moiety is selected from the groupconsisting of:

—O—(CH₂)_(m)—O—; —{NH—CHR—(CO)}_(m)—O—; —O—(CF₂)_(m)—O—; —(S)_(m)—;—O(CH₂CH₂O)_(m)—; —(OCH(CH₃)CH₂)_(m)O—;

Non-limiting examples of bridged macrocyclic module compounds includethe following:

wherein R¹ is CH₂CO₂(CH₂)₁₅CH₃;

wherein R^(o) is H, alkyl, or a lipophilic group; wherein R′ is anatural α-amino acid side chain; and wherein the structure

may be either benzene or cyclohexane. In one embodiment,

is benzene. In another embodiment,

is cyclohexane.

In another aspect are nanofilms comprising a plurality of a bridgedmacrocyclic module as defined herein. In one embodiment, the thicknessof the nanofilm composition is less than about 30 nanometers. In anotherembodiment, the thickness of the nanofilm composition is less than about6 nanometers. In another embodiment, the nanofilm is impermeable toviruses and larger species. In another embodiment, the nanofilm isimpermeable to immunoglobulin G and larger species. In anotherembodiment, the nanofilm is impermeable to albumin and larger species.In another embodiment, the nanofilm is impermeable to β2-Microglobulinand larger species. In another embodiment, the nanofilm is permeableonly to water and smaller species. In another embodiment, the nanofilmhas a molecular weight cut-off of 13 kDa. In another embodiment, thenanofilm has a molecular weight cut-off of 190 Da. In anotherembodiment, the nanofilm has a molecular weight cut-off of 100 Da. Inanother embodiment, the nanofilm has a molecular weight cut-off of 45Da. In another embodiment, the nanofilm has a molecular weight cut-offof 20 Da. In another embodiment, the nanofilm has high permeability forwater molecules and Na+, K+, and Cs+ in water. In another embodiment,the nanofilm has low permeability for glucose and urea. In anotherembodiment, the nanofilm has high permeability for water molecules andCl− in water. In another embodiment, the nanofilm has high permeabilityfor water molecules and K+ in water, and low permeability for Na+ inwater. In another embodiment, the nanofilm has high permeability forwater molecules and Na+ in water, and low permeability for K+ in water.In another embodiment, the nanofilm has low permeability for urea,creatinine, Li+, Ca2+, and Mg2+ in water. In another embodiment, thenanofilm has high permeability for Na+, K+, hydrogen phosphate, anddihydrogen phosphate in water. In another embodiment, the nanofilm hashigh permeability for Na+, K+, and glucose in water. In anotherembodiment, the nanofilm has low permeability for myoglobin, ovalbumin,and albumin in water. In another embodiment, the nanofilm has highpermeability for organic compounds and low permeability for water. Inanother embodiment, the nanofilm has low permeability for organiccompounds and high permeability for water. In another embodiment, thenanofilm has low permeability for water molecules and high permeabilityfor helium and hydrogen gases. In another embodiment is a nanofilmcomposition comprising at least two layers of a nanofilm defined herein.In another embodiment, the nanofilm composition comprises at least onespacing layer between any two of the nanofilm layers. In one embodiment,the spacing layer comprises a layer of a polymer, a gel, or inorganicparticles. In another embodiment, the nanofilm is deposited on asubstrate. In one embodiment, the substrate is porous. In anotherembodiment, the nanofilm is coupled to the substrate throughbiotin-strepavidin mediated interaction.

In another aspect is a method of filtration comprising using a nanofilmof the invention to separate components from fluid.

In another aspect is a method for making a bridged macrocyclic modulecompound of the invention, comprising: (a) providing a bridged programdirector compound of the structure:

wherein each J¹ is a functional group for coupling an adjacent synthon;and (b) reacting a synthon or a synthon multimer with said bridgedprogram director compound to form a bridged macrocyclic module compound.

In another aspect is a method for making a bridged macrocyclic modulecompound of the invention, comprising: (a) providing a macrocyclicmoiety compound, wherein the macrocyclic moiety compound contains from 4to 50 synthons; and (b) reacting a bridge moiety comprising at least twotermini with said macrocyclic moiety compound to form a bridgedmacrocyclic module compound.

This invention further includes the rational design of bridgedmacrocyclic module compounds that may be assembled into largerconstructions. Bridged macrocyclic module compounds may be used fromwhich hierarchical molecules approaching macroscopic dimension havingpredicted properties can be assembled. These bridged macrocyclic modulesmay have functional groups which couple to complementary functionalgroups on other bridged macrocyclic modules to create larger structures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows examples of surface pressure vs. area isotherms of ananofilm of Octamer IV pjs.

FIG. 2 shows examples of Brewster Angle Microscopy (BAM) data for ananofilm of Octamer IV pjs.

FIG. 3 shows the conformation of Octamer IV pjs after molecular dynamicssimulation.

FIG. 4A shows the conformation of Octamer IV pjs before molecularmechanics annealing routines.

FIG. 4B shows two views of the conformation of Octamer IV pjs aftermolecular mechanics annealing routines.

FIG. 5A shows the conformation of a molecule of Octamer IV pjs withoutthe bridge moiety before and after a molecular mechanics annealingroutine.

FIG. 5B shows the conformation of Octamer IV pjs before and after amolecular mechanics annealing routine.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The term “synthon” is used herein to indicate a monomeric molecular unitfrom which a macrocyclic module may be made; a macrocyclic module is aclosed ring of coupled synthons, for example, from 4 to 50, or moresynthons. Structures and syntheses of synthons and macrocyclic modulesare described in greater detail hereinbelow.

A macrocyclic module may be coupled with a molecular bridge (“bridgemoiety”), which comprises at least two termini, to form a bridgedmacrocyclic module. When part of a bridged macrocylic module, themacrocyclic module portion may be termed “macrocyclic moiety.”

The term “bridge moiety” is used herein to indicate a molecular bridge.The bridge moiety may have two termini, resulting in a bicyclicstructure when the two termini of the bridge moiety are coupled to themacrocyclic moiety. The bridge may have three or more termini, resultingin a polycyclic structure when at least three termini are coupled to themacrocyclic moiety. The bridge may have, for example, 3, 4, 5, 6, 8, 12,16, 32, 64 or more termini, at least two of which may couple to themacrocyclic moiety.

The terms “bridged macrocyclic module” and “bridged macrocyclic modulecompound” are used interchangeably herein and are used to indicate acompound comprising a macrocyclic moiety and a bridge moiety. The bridgemoiety comprises at least two termini, resulting in a bicyclic orpolycyclic structure when the two or greater termini are coupled to themacrocyclic moiety. In a “bridged macrocyclic module”, not all terminiof the bridge moiety must be coupled to the macrocyclic moiety, however,a minimum of two termini must be coupled to the macrocyclic moiety.Bridged macrocyclic modules may be coupled to each other to formmacromolecular structures, for example, a nanofilm.

The term “synthon multimer” is used herein to indicate a linear couplingof two or more synthons. For example, “synthon trimer” is used toindicate a molecule of the general formula: synthon-synthon-synthon.

The term “activated synthon multimer” is used herein to indicate asynthon multimer with functional groups for coupling the activatedsynthon multimer to additional synthons or to a bridged programdirector, preferably in a specific orientation.

The term “bridged program director” is used herein to indicate acompound comprising a bridge moiety and at least two synthons, whereinthe bridge moiety has 2 or more termini, and wherein each synthon iscoupled to a terminus of the bridge moiety, and wherein the compoundcomprises functional groups to permit the coupling of additionalsynthons or synthon multimers to the bridged program director,preferably in a specific orientation.

The term “component” is used herein to refer to the molecules used inproducing the bridged macrocyclic module compounds and 2-D and 3-Dhierarchical structures produced therefrom, e.g., synthons, bridgemoieties, macrocyclic moieties, macrocyclic modules, bridged macrocyclicmodules, and substrates.

As used herein, the terms “amphiphile” or “amphiphilic” refer to amolecule or species which exhibits both hydrophilic and lipophiliccharacter. In general, an amphiphile contains a lipophilic moiety and ahydrophilic moiety. The terms “lipophilic” and “hydrophobic” areinterchangeable as used herein. An amphiphile may form a Langmuir film.

Non-limiting examples of hydrophobic groups or moieties include loweralkyl groups, alkyl groups having 7, 8, 9, 10, 11, 12, or more carbonatoms, including alkyl groups with 14-30, or 30 or more carbon atoms,substituted alkyl groups, alkenyl groups, alkynyl groups, aryl groups,substituted aryl groups, saturated or unsaturated cyclic hydrocarbons,heteroaryl, heteroarylalkyl, heterocyclic, and corresponding substitutedgroups. A hydrophobic group may contain some hydrophilic groups orsubstituents insofar as the hydrophobic character of the group is notoutweighed. In further variations, a hydrophobic group may includesubstituted silicon atoms, and may include fluorine atoms. Thehydrophobic moieties may be linear, branched, or cyclic. Non-limitingexamples of preferred groups which may be coupled to a synthon ormacrocyclic module as a lipophilic group include alkyls, —CH═CH—R³,—C≡C—R³, —OC(O)—R³, —C(O)O—R³, —NHC(O)—R³, —C(O)NH—R³, —O—R³, and—CH₂COOR³, where R³ is 4-18C alkyl.

Non-limiting examples of hydrophilic groups or moieties includehydroxyl, methoxy, phenol, carboxylic acids and salts thereof, methyl,ethyl, and vinyl esters of carboxylic acids, amides, amino, cyano,isocyano, nitrile, ammonium salts, sulfonium salts, phosphonium salts,mono- and di-alkyl substituted amino groups, polypropyleneglycols,polyethylene glycols, epoxy groups, acrylates, sulfonamides, nitro,—OP(O)(OCH₂CH₂N⁺RRR)O⁻, guanidinium, aminate, acrylamide, pyridinium,piperidine, and combinations thereof, wherein each R is independentlyselected from H or alkyl. Further examples include polymethylene chainssubstituted with alcohol, carboxylate, acrylate, methacrylate, or

groups, where y is 1-6. Hydrophilic moieties may also include alkylchains having internal amino or substituted amino groups, for example,internal —NH—, —NC(O)R—, or —NC(O)CH═CH₂— groups, wherein R is H oralkyl. Hydrophilic moieties may also include polycaprolactones,polycaprolactone diols, poly(acetic acid)s, poly(vinyl acetates)s,poly(2-vinyl pyridine)s, cellulose esters, cellulose hydroxyl ethers,poly(L-lysine hydrobromide)s, poly(itaconic acid)s, poly(maleic acid)s,poly(styrenesulfonic acid)s, poly(aniline)s, or poly(vinyl phosphonicacid)s. A hydrophilic group may contain some hydrophobic groups orsubstituents insofar as the hydrophilic character of the group is notoutweighed.

As used herein, the terms “coupling” and “coupled” with respect tomolecular moieties or species, atoms, synthons, macrocyclic modules andbridged macrocyclic modules refers to their attachment or associationwith other molecular moieties or species, atoms, synthons, macrocyclicmodules and bridged macrocyclic modules. The attachment or associationmay be specific or non-specific, reversible or non-reversible, theresult of chemical reaction, or complexation or charge transfer. Thebonds formed by a coupling reaction are often covalent bonds, orpolar-covalent bonds, or mixed ionic-covalent bonds, and may sometimesbe Coulombic forces, ionic or electrostatic forces or interactions. Inpreferred embodiments, the coupling is covalent.

As used herein, the terms “R,” “R′,” “R″”, and “R′″” in a chemicalformula refer to a hydrogen or a functional group, each independentlyselected, unless stated otherwise. In some embodiments the functionalgroup may be an organic group. In some embodiments the functional groupmay be an alkyl group. In some embodiment, the functional group may be alipophilic group.

As used herein, the term “functional group” includes, but is not limitedto, chemical groups, biochemical groups, organic groups, inorganicgroups, organometallic groups, aryl groups, heteroaryl groups, cyclichydrocarbon groups, amino (—NH₂), hydroxyl (—OH), cyano (—C≡N), nitro(NO₂), carboxyl (—COOH), formyl (—CHO), keto (—CH₂C(O)CH₂—), alkenylalkynyl, (—C≡C—), and halo (F, Cl, Br and I) groups. In someembodiments, the functional group is an organic group.

As used herein, the term “alkyl” refers to a branched or unbranchedmonovalent hydrocarbon radical. An “n-mC” alkyl or “(nC-mC)alkyl” refersto all alkyl groups containing from n to m carbon atoms. For example, a1-4C alkyl refers to a methyl, ethyl, propyl, or butyl group. Allpossible isomers of an indicated alkyl are also included. Thus, propylincludes isopropyl, butyl includes n-butyl, isobutyl and t-butyl, and soon. An alkyl group with from 1-6 carbon atoms is referred to as “loweralkyl.” The term alkyl includes substituted alkyls.

As used herein, the term “substituted alkyl” refers to an alkyl groupwith an additional group or groups attached to any carbon of the alkylgroup. Substituent groups may include one or more functional groups suchas alkyl, lower alkyl, aryl, acyl, halogen, alkylhalo, hydroxy, amino,alkoxy, alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto,both saturated and unsaturated cyclic hydrocarbons, heterocycles, andother organic groups.

As used herein, the term “alkenyl” refers to any structure or moietyhaving the unsaturation C═C. As used herein, the term “alkynyl” refersto any structure or moiety having the unsaturation C≡C.

As used herein, the term “aryl” refers to an aromatic group which may bea single aromatic ring or multiple aromatic rings which are fusedtogether, coupled covalently, or coupled to a common group such as amethylene, ethylene, or carbonyl, and includes polynuclear ringstructures. An aromatic ring or rings may include substituted orunsubstituted phenyl, naphthyl, biphenyl, diphenylmethyl, andbenzophenone groups, among others. The term “aryl” includes substitutedaryls.

As used herein, the term “substituted aryl” refers to an aryl group withan additional group or groups attached to any carbon of the aryl group.Additional groups may include one or more functional groups such aslower alkyl, aryl, acyl, halogen, alkylhalos, hydroxy, amino, alkoxy,alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, thioether,heterocycles, both saturated and unsaturated cyclic hydrocarbons whichare fused to the aromatic ring(s), coupled covalently or coupled to acommon group such as a methylene or ethylene group, or a carbonylcoupling group such as in cyclohexyl phenyl ketone, and others.

As used herein, the term “heteroaryl” refers to an aromatic ring(s) inwhich one or more carbon atoms of the aromatic ring(s) are substitutedby a heteroatom such as nitrogen, oxygen, or sulfur. Heteroaryl refersto structures which may include a single aromatic ring, multiplearomatic rings, or one or more aromatic rings coupled to one or morenonaromatic rings. It includes structures having multiple rings, fusedor unfused, coupled covalently, or coupled to a common group such as amethylene or ethylene group, or coupled to a carbonyl as in phenylpyridyl ketone. As used herein, the term “heteroaryl” includes ringssuch as thiophene, pyridine, isoxazole, phthalimide, pyrazole, indole,furan, or benzo-fused analogues of these rings.

As used herein, the term “acyl” refers to a carbonyl substituent,—C(O)R⁶, where R⁶ is alkyl or substituted alkyl, aryl or substitutedaryl, which may be called an alkanoyl substituent when R⁶ is alkyl.

As used herein, the term “amino” refers to a group —NR⁴R⁵, where R⁴ andR⁵ may independently be hydrogen, lower alkyl, substituted lower alkyl,aryl, substituted aryl or acyl.

As used herein, the term “alkoxy” refers to an —OR⁷ group, where R⁷ isan alkyl, substituted lower alkyl, aryl, substituted aryl. Alkoxy groupsinclude, for example, methoxy, ethoxy, phenoxy, substituted phenoxy,benzyloxy, phenethyloxy, t-butoxy, and others.

As used herein, the term “thioether” refers to the general structureR⁸—S—R⁹ in which R⁸ and R⁹ are the same or different and may be alkyl,aryl or heterocyclic groups. The group —SH may also be referred to as“sulfhydryl” or “thiol” or “mercapto.”

As used herein, the term “saturated cyclic hydrocarbon” refers to ringstructures cyclopropyl, cyclobutyl, cyclopentyl groups, and others,including substituted groups. Substituents to saturated cyclichydrocarbons include substituting one or more carbon atoms of the ringwith a heteroatom such as nitrogen, oxygen, or sulfur. Saturated cyclichydrocarbons include bicyclic structures such as bicycloheptanes andbicyclooctanes, and multicyclic structures.

As used herein, the term “unsaturated cyclic hydrocarbon” refers to amonovalent nonaromatic group with at least one double bond, such ascyclopentenyl, cyclohexenyl, and others, including substituted groups.Substituents to unsaturated cyclic hydrocarbons include substituting oneor more carbon atoms of the ring with a heteroatom such as nitrogen,oxygen, or sulfur. Unsaturated cyclic hydrocarbons include bicyclicstructures such as bicycloheptenes and bicyclooctenes, and multicyclicstructures.

As used herein, the term “cyclic hydrocarbon” includes substituted andunsubstituted, saturated and unsaturated cyclic hydrocarbons, andmulticyclic structures.

As used herein, the term “heteroarylalkyl” refers to -alkyl-heteroaryl.

As used herein, the term “heterocyclic” refers to a monovalent saturatedor unsaturated nonaromatic group having a single ring or multiplecondensed rings having from 1-12 carbon atoms and from 1-4 heteroatomsselected from nitrogen, phosphorous, sulfur, or oxygen within the ring.Examples of heterocycles include tetrahydrofuran, morpholine,piperidine, pyrrolidine, and others.

As used herein, each chemical term described above expressly includesthe corresponding substituted group. For example, the term“heterocyclic” includes substituted heterocyclic groups.

As used herein, the term “activated acid” refers to a —C(O)X² moiety,where X² is a leaving group, in which the X² group is readily displacedby a nucleophile to form a covalent bond between the —C(O)— and thenucleophile. Examples of activated acids include acid chlorides, acidfluorides, p-nitrophenyl esters, pentafluorophenyl esters, andN-hydroxysuccinimide esters.

As used herein, the term “amino acid residue” refers to the productformed when a species comprising at least one amino (—NH₂) and at leastone carboxyl (—C(O)O—) group couples through either of its amino orcarboxyl groups with an atom or functional group of a synthon. Whicheverof the amino or carboxyl groups is not involved in the coupling may beblocked with a removable protective group.

Bridged Macrocyclic Module Compounds

In one aspect of the invention are compounds of the formula:

wherein each Q is an independently selected synthon, each L isindependently a bond or a linker molecule, n′ is from 4 to 50, andwherein the compound further comprises a bridge moiety having two ormore termini, wherein at least two of said two or more termini arecoupled to the compound. The termini of the bridge moiety may each beindependently coupled to either a synthon Q or a linker molecule L. Inone embodiment, the bridge moiety termini are coupled to the synthons Q.In another embodiment, the bridge moiety termini are coupled to thelinker molecules L. In a preferred embodiment, n is from 4 to 30, morepreferably 4 to 24, still more preferably 4 to 12.

In other embodiments are bridged macrocyclic module compounds of one ofthe following formulas:

wherein: A is a bridge moiety as defined herein; each Q¹ is anindependently selected synthon as defined herein; each Q² is anindependently selected synthon as defined herein; each Q³ is anindependently selected synthon as defined herein, and each L is anindependently selected bond or a linker molecule as defined herein.

In some embodiments, each Q¹ is the same synthon. In some embodiments,each Q² is the same synthon. In some embodiments, each Q³ is the samesynthon. In some embodiments, the synthons are directly coupled (L is abond). In other embodiments, the synthons are coupled to each otherthrough linker molecules.

In some embodiments, the bridge moiety may be connected to synthonsdirectly, for example:

wherein A is a bridge moiety, each Q is an independently selectedsynthon, each L is an independently selected linker molecule or a bond,and each n is independently from 1 to 24.

The bridge moiety may also be connected to the synthons through a linkermolecule, for example:

In other embodiments, the bridge moiety may be connected to a linkerbetween synthons, or to combinations of synthons and linkers, forexample:

These bridged macrocyclic modules may comprise functional groups whichcouple to complementary functional groups on other bridged macrocyclicmodules to create larger structures. These bridged macrocyclic modulesmay comprise functional groups for coupling the module to a substrate.

Exemplary, non-limiting, bridged macrocyclic module compounds includethe following:

wherein A is a bridge moiety comprising one or more of the followingstructural units:

wherein each R^(o) comprises H, an alkyl group, or a lipophilic group;wherein R′ comprises a natural α-amino acid side chain; and wherein X′is H or a functional group for coupling the bridged macrocyclic moduleto another bridged macrocyclic module or to a substrate.

Additional non-limiting examples of bridged macrocyclic module compoundsmay be found in the Examples and Claims.

Bridged Program Directors

In another aspect, this invention relates to bridged program directorcompounds, and compounds useful in the synthesis of bridged programdirector compounds. A bridged program director is a discrete productmolecule which has built-in directionality to control coupling reactionsof the terminal synthons to additional synthons.

In one aspect of the invention are compounds of the formula:

wherein A is a bridge moiety as defined herein; each Q^(n) is anindependently selected synthon as defined herein; and n is from 2 to 30.These compounds may be useful in the synthesis of bridged programdirector compounds.

In another aspect of the invention are bridged program directorcompounds of one of the following formula:

wherein A is a bridge moiety as defined herein; each Q¹ is anindependently selected synthon as defined herein; each J¹, if present,is an independently selected functional group for coupling one or moreadditional synthons; each Q², if present, is an independently selectedsynthon as defined herein; each Q³, if present, is an independentlyselected synthon as defined herein; and n is from 2 to 30.

In some embodiments, A is coupled to the synthons Q¹ directly. In otherembodiments, A is coupled to Q¹ through a linker molecule. In someembodiments, the synthons are coupled directly to each other (e.g., Q¹is coupled directly to Q², Q² is coupled directly to Q³). In otherembodiments, the synthons may be coupled through linker molecules (e.g.,Q¹ is coupled to Q² through a linker molecule, Q² is coupled to Q³through a linker molecule). In some embodiments, some sythons may beconnected through linker molecules, while other synthons are directlycoupled.

In a preferred embodiment, n is from 2 to 5. In a more preferredembodiment, n is from 2 to 3.

The use of bridged program directors may be advantageous in thesynthesis of bridged macrocyclic modules. The use of bridged programdirectors may allow for convergent synthesis methods of some bridgedmacrocyclic modules, which may result in fewer synthesis steps and, ingeneral, higher yields. Examples of convergent synthesis methods may befound hereinbelow and in, for example, Example 4. In some embodiments,the functional groups on the synthons may be tailored to only bind to aparticular additional synthon in a particular orientation. Further, thepositioning of functional groups on the synthons may be useful inlimiting side reactions.

Synthons

A general formula for a cyclic or an acyclic synthon may be written:

wherein T is a cyclic or an acyclic core synthon having attachedfunctional groups J′, X′, and Z; each J′ is an independently selectedfunctional group for coupling the synthon to one or more other synthonsor to a bridge moiety, wherein n″ is at least 2; each X′ isindependently H or a functional group for coupling the synthon toanother bridged macrocyclic module or to a substrate, wherein m″ is 0,1, or greater than 1; and each Z is independently either H, a lipophilicgroup, or a hydrophilic group, wherein p″ is 0, 1, or greater than 1.Functional groups J′ and X′ may be selected, for example, from thoselisted in Tables 6-8. Preferred lipophilic groups Z include, forexample, alkyls, —CH═CH—R³, —C≡C—R³, —OC(O)—R³, —C(O)O—R³, —NHC(O)—R³,—C(O)NH—R³, —CH₂COOR³, and —O—R³, where R³ is 4-18C alkyl.

In one embodiment, the X′ group on a synthon is H. In anotherembodiment, the X′ group on a synthon is a functional group. In someembodiments, a synthon may be substantially one isomeric configuration,for example, a single enantiomer.

Cyclic Synthons

As used herein, the term “cyclic synthon” refers to a synthon having oneor more ring structures. Examples of ring structures include aryl,heteroaryl, and cyclic hydrocarbon structures including bicyclic ringstructures and multicyclic ring structures. Examples of core cyclicsynthons include benzene, cyclohexadiene, cyclopentadiene, naphthalene,anthracene, phenylene, phenanthracene, pyrene, triphenylene,phenanthrene, pyridine, pyrimidine, pyridazine, biphenyl, bipyridyl,cyclohexane, cyclohexene, decalin, piperidine, pyrrolidine, morpholine,piperazine, pyrazolidine, quinuclidine, tetrahydropyran, dioxane,tetrahydrothiophene, tetrahydrofuran, pyrrole, cyclopentane,cyclopentene, triptycene, adamantane, bicyclo[2.2.1]heptane,bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane, bicyclo[2.2.2]octene,bicyclo[3.3.0]octane, bicyclo[3.3.0]octene, bicyclo[3.3.1]nonane,bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane, bicyclo[3.2.2]nonene,bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, and spiro[4.4]nonane. A core synthoncomprises all isomers or arrangements of coupling the core synthon toother synthons. For example, the core synthon benzene includes synthonssuch as 1,2- and 1,3-substituted benzenes, where the linkages betweensynthons are formed at the 1,2- and 1,3-positions of the benzene ring,respectively. For example, the core synthon benzene includes1,3-substituted synthons such as

where L is a linkage between synthons and the 2,4,5,6 positions of thebenzene ring may also have substituents. A condensed linkage betweensynthons involves a direct coupling between a ring atom of one cyclicsynthon to a ring atom of another cyclic synthon, for example, wheresynthons Q-X and Q-X couple to form Q-Q, where Q is a cyclic synthon andX is halogen; as for example when Q is phenyl resulting in the condensedlinkage

Examples of synthons and their syntheses are further described in U.S.patent application Ser. Nos. 10/071,377 and 10/226,400 filed Feb. 7,2002 and Aug. 23, 2002, respectively, and in PCT Application No.PCT/US03/03830, filed Feb. 7, 2003.

Acyclic Synthons

Examples of formulas representing acyclic synthons are shown in Table 1.

TABLE 1 Examples of acyclic synthons

J and X′ may be a functional group selected, for example, from thosefound in Tables 6-8. Z may be any lipophilic group. Preferred lipophilicgroups include alkyls, —CH═CH—R³, —C≡C—R³, —OC(O)—R³, —C(O)O—R³,—NHC(O)—R³, —C(O)NH—R³, —CH₂COOR³, and —O—R³, where R³ is 4-18C alkyl.The synthons may further comprise functional groups for coupling with abridge moiety, nonlimiting examples of suitable functional groups andlinkages formed may be found in Tables 6-8.

Non-limiting examples of acyclic core synthons are shown in Table 2.

TABLE 2 Examples of acyclic core synthons —OCH₂CH₂——(CH₂)_(n)C≡C(CH₂)_(n)—

—(CH₂)_(n)— —C(O)O(CH₂)_(n)— —(CH₂)_(n)C(O)NR^(o)— —S_(m)— (m = 2-14)—(CH₂)_(n)SiMe₂(CH₂)_(n)— —(CH₂)_(n)NR^(o)(CH₂)_(n)— —(CH₂)_(n)CH(OH)—

In Table 2, R^(o) is H or a lipophilic group, and n is 1-22.

The examples of acyclic synthons in Table 2 may further comprise thegroups X′, J and Z as described above, as well as functional groups forcoupling to the bridge moiety.

Suitable acyclic synthons may be purchased from, for example, AldrichChemical Company (St. Louis, Mo.). Alternatively, one skilled in the artmay synthesize appropriate acyclic synthons.

Preferred synthons for the bridged macrocyclic module compounds include:

wherein each X′ are independently H or a functional group for couplingthe synthon to another bridged macrocyclic moiety (“cross-linkinggroups”), each J are independently selected functional groups forcoupling the synthons to additional synthons within the same bridgedmacrocyclic moiety, and X₁ are functional groups for coupling thesynthon to a bridge moiety. In a preferred embodiment, each X′ within asynthon is the same. In another preferred embodiment, each J within asynthon is the same.

Particularly preferred synthons for a bridged macrocyclic module include

wherein each X′ are independently H or a functional group for couplingthe synthon to another bridged macrocyclic moiety (“cross-linkinggroups”), each J are independently selected functional groups forcoupling the synthons to additional synthons within the same bridgedmacrocyclic moiety, and X₁ are functional groups for coupling thesynthon to a bridge moiety. In a preferred embodiment, each X′ within asynthon is the same. In another preferred embodiment, each J within asynthon is the same.

Macrocyclic Modules

A macrocyclic module is a closed ring of 4-50 coupled synthons. Thesynthons may be coupled using, for example, the functional groups inTables 6-8. A macrocyclic module may be coupled to a bridge moietyhaving at least two termini to produce a bridged macrocyclic module, asfurther described herein.

The preparation of macrocyclic modules beginning with a set of synthonsis described in U.S. patent application Ser. Nos. 10/071,377 and10/226,400, and in the PCT Application PCT/US03/03830, incorporated byreference herein in their entirety. The assembly of molecular buildingblocks, beginning with a set of synthons assembled to make macrocyclicmodules, which, in turn, are combined to form a nanofilm are describedin U.S. Ser. No. 60/383,236, filed May 22, 2002, and in U.S. patentapplication Ser. No. 10/359,894, filed Feb. 7, 2003, incorporated byreference herein in their entirety. Examples and syntheses of synthons,macrocyclic modules, and amphiphilic macrocyclic modules are furtherdescribed hereinbelow.

Examples of macrocyclic modules which may be modified with a bridgemoiety to produce a bridged macrocyclic module include those shown inTable 3.

TABLE 3 Examples of macrocyclic modules MODULE STRUCTURE Hexamer 1a

Hexamer 1dh

Hexamer 3j- amine

Hexamer 1jh-AC

Hexamer 1jh

Hexamer 2j- amine/ester

Hexamer 1dh- acryl

Octamer 5jh- aspartic

Octamer 4jh- acryl

An individual macrocyclic module may include a pore in its structure,which has a particular size depending on the conformation and state ofthe module. Addition of a bridge moiety to the module to form a bridgedmacrocyclic module may result in pores with different sizes andproperties.

Macrocyclic modules and bridged macrocyclic modules may have varyingdegrees of flexibility in their structures. In general, addition of abridge moiety to a macrocyclic module decreases the flexibility of themodule. Increased flexibility of the macrocyclic modules or bridgedmacrocyclic modules may allow the modules to more easily form linkageswith other modules by coupling reactions. Flexibility of a macrocyclicmodule or bridged macrocyclic module may also play a role in regulatingpassage of species through the pore of the module. For example,flexibility may affect the dimension of the pore of an individual modulesince various conformations may be available to the structure. Forexample, a module may have a certain pore dimension in one conformationwhen one group of substituents are located at the pore, and have adifferent pore dimension in a different conformation when a differentgroup of substituents are located at the pore. For example, the “onegroup” of substituents located at the pore may be three alkoxy groupsarranged in one regioisomer, while the “different group” of substituentsmay be two alkoxy groups arranged in another regioisomer. The effect ofthe “one group” of substituents located at the pore and the “differentgroup” of substituents located at the pore is to provide a bridgedmacrocyclic module composition which may regulate transport andfiltration, in conjunction with other regulating factors.

Bridge Moieties

Examples of bridge moieties include polymers, biopolymers, chainstructures, and various other chemical groups and structures. Bridgemoieties may also be comprised of multimeric cores.

Examples of polymers and biopolymers which may be used as a bridgemoiety include poly(maleic anhydrides), a copolymer of maleic anhydride,poly(ethylene-co-maleic anhydride), poly(maleic anhydride-co-alphaolefin), polyacrylates, a polymer or copolymer having acrylate sidegroups, a polymer or copolymer having oxacyclopropane side groups,polyethyleneimides, polyetherimides, polyethylene oxides, polypropyleneoxides, polystyrenes, poly(vinyl acetate)s, polytetrafluoroethylenes,polyolefins, polyethylenes, polypropylenes, ethylene-propylenecopolymers, polyisoprenes, neopropenes, polyanilines, polyacetylenes,polyvinylchlorides, polyvinylidene chlorides, polyvinylidene fluorides,polyvinylalcohols, polyurethanes, polyamides, polyimides, polysulfones,polyethersulfones, polysulfonamides, polysulfoxides, polyglycolic acids,polyacrylamides, polyvinylalcohols, polyesters, polyester ionomers,polyethylene terephthalates, polybutylene terephthalates,polycarbonates, polysorbates, polylysines, polypeptides, poly(aminoacids), polyvinylpyrrolidones, polylactic acids, gels, hydrogels,carbohydrates, polysaccharides, agarose, amylose, amylopectin, glycogen,dextran, cellulose, cellulose acetates, chitin, chitosan, peptidoglycan,and glycosaminoglycan. Further examples include amino-branched,amino-substituted, and amino-terminal derivatives of the precedingexample polymers. Further examples include polynucleotides, synthetic ornaturally-occurring polynucleotides, for example, poly(T) and poly(A),nucleic acids, as well as proteoglycans, glycoproteins, and glycolipids.

Examples of polymerizable monomers which can be used to form thepolymeric bridge moiety include vinyl halide compounds such as vinylchloride; vinylidene monomers such as vinylidene chloride; unsaturatedcarboxylic acids such as acrylic acid, methacrylic acid, maleic acid,itaconic acid, and salts thereof; acrylates such as methyl acrylate,ethyl acrylate, butyl acrylate, octyl acrylate, methoxyethyl acrylate,phenyl acrylate and cyclohexyl acrylate; methacrylates such as methylmethacrylate, ethyl methacrylate, butyl methacrylate, octylmethacrylate, phenyl methacrylate and cyclohexyl methacrylate;unsaturated ketones such as methyl vinyl ketone, ethyl vinyl ketone,phenyl vinyl ketone, methyl isobutenyl ketone and methyl isopropenylketone; vinyl esters such as vinyl formate, vinyl acetate, vinylpropionate, vinyl butyrate, vinyl benzoate, vinyl monochloroacetate,vinyl dichloroacetate, vinyl trichloroacetate, vinyl monofluoroacetate,vinyl difluoroacetate and vinyl trifluoroacetate; vinyl ethers such asmethyl vinyl ether and ethyl vinyl ether; acrylamide and alkylsubstituted compounds thereof; acid compounds containing a vinyl groupand salts, anhydrides and derivatives thereof such as vinylsulfonicacid, allylsulfonic acid, methallylsulfonic acid, styrenesulfonic acid,2-acrylamido-2-methylpropanesulfonic acid, sulfopropyl methacrylate,vinylstearic acid and vinylsulfinic acid; styrene or alkyl- orhalogen-substituted compounds thereof such as styrene, methylstyrene andchlorostyrene; allyl alcohol or esters or ethers thereof; vinylimidessuch as N-vinylphthalimide and N-vinylsuccinoimide; basic vinylcompounds such as-vinylpyridine, vinylimidazole, dimethylaminoethylmethacrylate, N-vinylpyrrolidone, N-vinylcarbazole and vinylpyridine;unsaturated aldehydes such as acrolein and methacrolein; andcross-linking vinyl compounds such as glycidyl methacrylate,N-methylolacrylamide, hydroxyethyl methacrylate, triallyl isocyanurate,triallyl cyanurate, divinylbenzene, ethylene glycol di(meth)acrylate,diethylene glycol di(meth)acrylate, triethylene glycol di(meth)acrylate,trimethylolpropane tri(meth)acrylate, and methylene bisacrylamide.

Examples of chain structures which may be used as a bridge moietyinclude chains containing one or more aryl groups, alkynyl groups,alkenyl groups, alkyl groups, ester groups, ether groups, amino acidresidues, sulfur atoms, substituted amine groups, alcohol groups, orsilicon atoms.

Non-limiting examples of multimeric cores include silsesquioxanes,dendrimers, and porphyrins. Silsesquioxanes (POSS) multimeric cores maybe obtained from, for example, Aldrich Chemical Company (Milwaukee,Wis.) and Hybrid Plastics (Fountain Valley, Calif.). Dendrimers anddendrimeric macromolecules may be obtained from, for example, AldrichChemical Company (Milwaukee, Wis.) and Dendritech, Inc. (Midland,Mich.). Porphyrins may be obtained from, for example, Aldrich ChemicalCompany (Milwaukee, Wis.) and Frontier Scientific Inc. (Logan, Utah).

Non-limiting examples of bridge moieties are shown in Table 4.

TABLE 4 Examples of bridge moieties

—O(CH₂)_(n)—O— —{NH—CHR—(CO)}_(n)—O— —O(CF₂)_(n)—O— —(S)_(m)— m = 2-14—O(CH₂CH₂O)_(n)— —(OCH(CH₃)CH₂)_(n)O—

In Table 4, n is 2-4.

Further example of bridge moieties include:

wherein X′ is independently H or a functional group for linking toanother bridged macrocyclic moiety or to other moieties, such ashydrophobic groups, biocompatibility groups, etc; and wherein R may be afunctional group for coupling the bridge moiety to a macrocyclic module,or may be an additional functional group, such as hydrogen, an activatedacid, —OH, —C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl, —NR′R′, —NR′R′R′⁺, —MgX,—Li, —OLi, —OK, —ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃,—NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂, —CH═CHR′, —CH═CR′₂,4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂, —C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

—P(O)(OH)(OX), or —P(═O)(O⁻)O(CH₂)_(s)NR′₃ ⁺;wherein R′ are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4.

In further variations, the bridge moiety may be star-shaped ordendrimeric. In these variations, a synthon is coupled to each of thetermini.

Preferred bridge moieties include

wherein X is a functional group for coupling the bridge moiety to asurface, and wherein R′ is a natural amino acid side chain.

Particularly preferred bridge moieties for the bridged macrocyclicmodules of the invention include

In one variation, the termini of a bridge moiety may be coupled tosynthons to form a bridged program director. The synthons coupled to abridged program director may have protection groups for directing theaddition of other synthons to the bridged program director. The bridgedprogram director has a built-in directionality which may control thecoupling of the synthons at the termini to other synthons.

In another variation, the bridge moiety may also be directly coupled toa macrocyclic module to form a bridged macrocyclic module.

Other variations on methods for producing bridged macrocyclic modulesfrom bridge moieties will be apparent from the methods and Examplesherein, as well as further variations apparent to those of skill in theart.

The bridge moiety may provide a variety of functions. For example, thebridge moiety may comprise functional groups for attachment of themodule to a surface. The chemical and steric properties of the bridgemoiety may affect transport modification and selectivity, throughchemical or electronic interaction with and/or steric inhibition ofcomponents of a fluid in contact with the bridged macrocyclic module.For example, adding steric bulk to the bridge might be expected todecrease effective pore size of the module. For example, use of apolypropylene glycol (PPG) bridge moiety versus a polyethylene glycol(PEG) bridge may result in a dramatically different solute sizeexclusion.

The bridge moiety may further comprise groups which help withbiocompatibility. In one example, PEG may be used as a bridge moiety,which may help to decrease plasma buildup. The bridge moiety may providethe module with amphiphilic character, which may, for example,facilitate the orientation of the modules on a surface to form ananofilm.

The bridge moieties may help to constrain the geometry of the molecule.For example, in comparison with a similar macrocylic module which doesnot contain a bridge moiety, the structure of the bridged macrocyclicmoiety may be more rigid.

Polymerizable Groups of Bridge Moieties

A bridge moiety may contain a polymerizable group, which may be usefulin the production of 2-D and 3-D arrays of bridged macrocyclic modules.For example, a polymerizable group may participate in anionic, cationic,radical, condensation, ring opening or other types of polymerizations. Apolymerizable group may have more than one polymerizable moiety, andseveral polymerizable groups may be attached to one bridge moiety.Examples of polymerization groups include those shown in Table 5.

TABLE 5 Examples of polymerizable groups of bridge and moieties

A non-limiting example of a bridged macrocyclic moiety with apolymerization groups is the structure:

Various monomers may be mixed with a bridged macrocyclic module whichcontains a polymerizable group and a polymerization may be performed,coupling the bridged macrocylic modules together (see examples inSchemes 1 and 2). In Scheme 1, R indicates H or a lipophilic group.

Formation of 2-D versus 3-D structures may be controlled by the type ofpolymerizable group added, as well as by the relative ratios of monomerto bridged macrocyclic modules. For example, in the schematic above,relatively higher amounts, or ratios, of 2-methyl-acrylic acid methylester versus the bridged macrocyclic module may favor formation of a2-dimensional structure rather than a 3-dimensional structure.

Network structures with higher 3D cross-linking density may also beachieved by using bridged macrocyclic modules or monomers which aremulti-functional. For example, an acrylamide polymerization scheme belowillustrates a bridged macrocyclic module with multiple polymerizationgroups and an analogous multi-functional cross-linking monomer:

In addition to the multi-component schemes above, a bridged macrocyclicmodule could also be self-polymerized. Such a polymerization would forma more discrete network of bridged macrocyclic modules with narrowerpore size distribution.

Coupling Methods

A variety of coupling schemes may be used in coupling the variouscomponents of the compositions of the invention. For example, synthonsmay possess functional groups for coupling of the synthon to othersynthons on the same or different bridged macrocyclic module, forcoupling of the synthon to bridge moieties on the same or differentbridged macrocyclic module, or for coupling of the synthon to asubstrate. Bridge moieties may possess functional groups for coupling ofthe bridge moiety to synthons, to other bridge moieties, or to asubstrate.

In one type of coupling, the linkage may be the product of the directcoupling of one functional group from each component. For example, ahydroxyl group of a first synthon may couple with an acid group or acidhalide group of a second synthon to form an ester linkage between thetwo synthons. Another example is an imine linkage, —CH═N—, resultingfrom the reaction of an aldehyde, —CH═O, on one synthon with an amine,—NH2, on another synthon. Non-limiting examples of suitable linkagesbetween, e.g., bridged macrocyclic modules, are shown in Table 6.

TABLE 6 Examples of functional groups and linkages formed FunctionalGroup A Functional Group B Linkage Formed —NH₂ —C(O)H —N═CH— —NH₂ —CO₂H—NHC(O)— —NHR —CO₂H —NRC(O)— —OH —CO₂H —OC(O)— —X —ONa —O— —SH —SH —S—S——X —(NR)Li —NR— —X —SNa —S— —X —NHR —NR— module-X

—X —CH₂CuLi —CH₂— —X —(CRR)_(n=1-6)CuLi —(CRR)_(n)— module-X module-Xmodule-module —CH₂X —CH₂X —CH₂CH₂— —ONa —C(O)OR —C(O)O— —SNa —C(O)OR—C(O)S— —X —C≡CH —C≡C— —C≡CH —C≡CH —C≡C—C≡C— —MgX —C(O)H —CH(OH)—Module-NH₂

Module-MgX

module-X

—C(O)H —C(O)H —HC═CH— (CH₃)₂C═CH-module module-C(O)Cl

—N═C═O —NH₂ —NHC(O)NH— —N═C═O HO— —NHC(O)O— —C(O)H —NHNH₂ —CH═N—NH— —OH—OC(O)X —OC(O)O— (CH₃)₂C═CH-module Module-SH

(CH₃)₂CHC(O)O-module module-CH(O)

module-CH₂C(O)OH module-CH₂C(O)OH

R₂SiH-module

—OP(O)(OH)₂ —OH —OP(O)(OH)O—

—NH₂

In Table 6, R independently represents a hydrogen or an alkyl group, andX is halogen or other good leaving group. It is to be understood thatthe functional groups and resulting linkages in Table 6 are not limitedto coupling modules to each other, but may also be used to couple othervarious components, for example, coupling a synthon to a substrate,coupling a synthon to a bridge moiety, etc.

In one variation, the functional group may be added to a component afterinitial preparation of that component. For example, a bridgedmacrocyclic module may have functional groups for coupling to otherbridged macrocyclic modules wherein the functional groups are coupled tothe bridged macrocyclic module after initial preparation of the closedring of the module. For example, an amine linkage between the synthonsof a bridged macrocyclic module may be substituted with one of variousfunctional groups to produce a substituted linkage. Examples of suchlinkages between synthons of a bridged macrocyclic module havingfunctional groups for coupling other bridged macrocyclic modules areshown in Table 7.

TABLE 7 Examples of macrocyclic module linkages Macrocyclic ModuleLinkage Reagent Substituted Linkage

In Table 7, X is halogen, and Q represents a synthon in a bridgedmacrocyclic module.

Referring to Table 7, the substituted linkage of a bridged macrocyclicmodule may couple to a substituted linkage of another module. In somevariations, the coupling of these linkages is done by initiating 2+2cycloaddition. For example, acrylamide linkages may couple to produce

by 2+2 cycloaddition. In other variations, coupling of these reactivesubstituted linkages may be initiated by other chemical, thermal,photochemical, electrochemical, and irradiative methods to provide avariety of coupled structures.

It is to be understood that the functional groups and substitutedlinkages formed included in Table 7 may also be used to couple othercomponents.

The functional groups used to form linkages between components may beseparated from the component by a spacer. A spacer can be any atom orgroup of atoms which couples the functional group to the component, anddoes not interfere with the linkage-forming reaction. A spacer is partof the functional group, and becomes part of the linkage betweencomponents. An example of a spacer is a polymethylene group, —(CH₂)n-,where n is 1-6. The spacer may be said to extend the linkage between thecomponents. Other examples of spacer groups are alkylene, aryl, acyl,alkoxy, saturated or unsaturated cyclic hydrocarbon, heteroaryl,heteroarylalkyl, heterocyclic, and corresponding substituted groups.Further examples of spacer groups are polymer, copolymer, or oligomerchains, for example, polyethylene oxides, polypropylene oxides,polysaccharides, polylysines, polypeptides, poly(amino acids),polyvinylpyrrolidones, polyesters, polyacrylates, polyamines,polyimines, polystyrenes, poly(vinyl acetate)s,polytetrafluoroethylenes, polyisoprenes, neopropene, polycarbonate,polyvinylchlorides, polyvinylidene fluorides, polyvinylalcohols,polyurethanes, polyamides, polyimides, polysulfones, polyethersulfones,polysulfonamides, polysulfoxides, and copolymers thereof. Examples ofpolymer chain spacer structures include linear, branched, comb anddendrimeric polymers, random and block copolymers, homo- andheteropolymers, flexible and rigid chains. The spacer may be any groupwhich does not interfere with formation of the linkage. A spacer groupmay be substantially longer or shorter than the functional group towhich it is attached.

Coupling of components to each other may occur through coupling offunctional groups of the components to linker molecules. The functionalgroups involved may be, for example, those exemplified in Table 8. Forexample, modules may couple to at least one other module through alinker molecule. A linker molecule is a discrete molecular species usedto couple at least two components. Each module may have 1 to 30 or morefunctional groups which may couple to a linker molecule. Linkermolecules may have 1 to 20 or more functional groups which may coupleto, for example, a module.

In one variation, a linker molecule has at least two functional groups,each of which can couple to a module and/or other component. In thesevariations, linker molecules may include a variety of functional groupsfor coupling modules and/or other components. Non-limiting examples offunctional groups of modules and linker molecules are illustrated inTable 8.

TABLE 8 Examples of functional groups of modules and linker moleculesFunctional Functional Group of Group of Module A Module B LinkerMolecule Linkage —NHR or —NH₂ —NHR or —NH₂

—NHR or —NH₂ —NHR or —NH₂

—NHR or —NH₂ —NHR or —NH₂

—NHR or —NH₂ —NHR or —NH₂

—OH —OH

—OH —OH

—OH —OH (RO)₂Br′B(OR)₂ —O(HO)Br′B(OH)O— —NHR or —NHR or (RO)₂Br′B(OR)₂—NH(HO)Br′B(OH)NH— —NH₂ —NH₂ —OH —OH X—(CH₂)_(n)—X —O—(CH₂)_(n)—O— —OH—OH ClC(O)—(CH₂)_(n)—C(O)Cl

—NHR or —NH₂ —NHR or —NH₂

—NHR or —NH₂ —NHR or —NH₂

—OH —OH

—OCH₂CH(OH)CH₂O— —OH —NH₂

—OCH₂CH(OH)CH₂NH— —NH₂ —NH₂

—NHCH₂CH(OH)CH₂NH— —NRH —NRH

—NHCH₂CH(OH)CH₂NR—

In Table 8, n is 1-6, m is 1-10, R is —CH₃ or —H, R′ is —(CH₂)_(n)— orphenyl, R″ is —(CH₂)—, polyethylene glycol (PEG), or polypropyleneglycol (PPG), and X is Br, Cl, I, or other good leaving groups which areorganic groups containing atoms selected from the group of carbon,oxygen, nitrogen, halogen, silicon, phosphorous, sulfur, and hydrogen. Amodule may have a combination of the various functional groupsexemplified in Table 8. It is to be understood that the functionalgroups and linkers included in Table 8 may also be used to link othercomponents together. Preferred linkers include DEM and ethylene diamine.Further examples of suitable linkers are found in the Examples.

Methods of initiating coupling of the components to linker moleculesinclude chemical, thermal, photochemical, electrochemical, andirradiative methods.

Other methods for linking the components will be apparent to one ofskill in the art.

Methods for Preparing Bridged Program Directors from Bridge Moieties

Scheme 4 illustrates an example of a general scheme for preparingbridged program director compounds.

In Scheme 4, A is a bridge moiety which provides at least two termini,each Q¹ is an independently selected synthon as defined herein, and eachJ is an independently selected functional group for further coupling toan independently selected synthon Q². Synthons Q² may be coupled tosynthons Q¹ directly, or through linkers. Examples of suitablefunctional groups and resulting linkages may be found in Tables 6-8.

The bridge moiety termini may comprise functional groups for couplingsynthons Q¹ to the termini. Alternatively, suitable functional groupsmay be added to the termini for coupling to the synthons. The synthonsmay be directly coupled to the termini, or may be coupled through linkermolecules. Examples of suitable functional groups and resulting linkagesare found in Tables 6-8. The termini may independently includeprotection groups.

In Scheme 4, synthons Q¹ are shown having functional groups J attached,before coupling to bridge moiety A. In an alternate scheme, synthons Q¹are coupled to bridge moiety A, and then synthons Q¹ are subsequentlyderivatized with functional groups J for coupling of additional synthonsQ².

Non-limiting examples for preparation of bridged program directorcompounds (compounds 4 and 5) may be found in Example 1. Additionalschemes and syntheses for preparing bridged program directors will beapparent to those of skill in the art.

Methods for Preparation of Activated Synthon Multimer Compounds:

Activated synthon multimers may be prepared by concerted and/orstep-wise methods. A general scheme for a concerted-approach is shown inScheme 5.

A non-limiting example for preparation of an activated synthon multimerA according to a concerted-approach may be found in Scheme 18 of Example4. Activated synthon multimers may also be prepared by step-wisemethods. Additional schemes and syntheses for preparing activatedsynthon multimer compounds will be apparent to those of skill in theart.

Methods for Preparing Bridged Macrocyclic Module Compounds

Various coupling schemes for preparing bridged macrocyclic modulecompounds may be used. A bridged macrocyclic module may be made via astepwise methodology, a pseudo-concerted approach, convergent methods,or a variety of other synthetic schemes that will be apparent to oneskilled in the art.

Pseudoconcerted Methods

A bridged macrocyclic module may be prepared by pseudoconcerted methods.For example, a bridged macrocyclic module may be prepared from a bridgedprogram director compound according to Scheme 6.

In Scheme 6, A is a bridge moiety, L are optional linker molecules, eachQ¹ is an independently selected synthon, each Q² is an independentlyselected synthon, and each Q³ is an independently selected synthon.Complementary functional groups present on synthons Q² and Q³ may reactto form the bicyclic bridged macrocyclic module. Q² and Q³ may bedirectly coupled or may be coupled through linker molecules. Examples ofsuitable functional groups, linkers and the resulting linkages forcoupling Q² to Q³ are found in Tables 6-8.

Non-limiting examples of the synthesis of bridged macrocyclic modulesaccording to this method may be found in Examples 1 and 2. It is to beunderstood that this approach may also be used for synthesizingpolycyclic bridged macarocyclic modules, wherein the bridge moietycomprises more than two termini. For example, Scheme 7 shows a generalapproach for the synthesis of a bridged macrocyclic module from abridged program director, wherein the bridged program director comprisesthree termini.

A specific example of a pseudo-concerted synthesis of a bridgedmacrocyclic module having a bridge moiety with four termini is shown inExample 5. Other variations on this method will be apparent to those ofskill in the art.

Stepwise Methods

In another variation, a bridged macrocyclic module may be synthesized bystepwise ring closure using a bridge moiety with appropriate functionalgroups, as shown in Scheme 8:

In Scheme 8, each synthon may be coupled directly to its adjacentsynthons, or may also be linked through a linker molecule. Examples ofsuitable functional groups, linkers and the resulting linkages are foundin Tables 6-8. A non-limiting example of the synthesis of a bridgedmacrocyclic module according to this method may be found in Example 3.

This approach may be of particular use in synthesizing asymmetricalbridged macrocyclic modules, although other methods may also be useful.It is to be understood that the method shown in Scheme 8 may also beused to produce a symmetrical bridged macrocyclic module. Furthervariations on this approach will be apparent to those of skill in theart.

Convergent Methods

A bridged macrocyclic module may also be prepared by convergent methods.For example, a bridged macrocyclic module is prepared from a bridgedprogram director and an activated synthon trimer according to Scheme 9.

In Scheme 9, an activated synthon multimer is reacted with a bridgedprogram director containing appropriate reactive functional groups,forming the bridged macrocylic module.

Schemes 10 and 11 illustrate a specific example of a synthesis of thistype.

Details of this synthesis are found in Example 4. The final assembly ofthe bridged macrocyclic module may be performed in situ. In separateflasks, the di-Boc protected synthon trimer is deprotected and atetra-acid bridged program director is activated, and immediatelythereafter, mixed together in a new flask to generate the desiredproduct as shown in Scheme 11.

The orientation of the functional groups on the bridged program directorhelp to limit the reaction products to the desired product. For example,in Scheme 11 above, the ortho-position of the acid chloride functionalgroups on synthons of the bridged program director limit the possibleorientations in which the activated synthon trimer may bind. Forexample, positioning of the acid chloride functional groups on themeta-position of the synthon in the bridged program director may resultin increased side reactions and possible polymer products.

Direct Synthesis from Macrocyclic Modules

Another method for the synthesis of bridged macrocyclic module compoundsis by linking a bridge moiety to a macrocyclic module. Scheme 12illustrates an example of this type of synthesis:

In Scheme 12, the bridge moiety A may be directly coupled to thesynthons Q¹, or may be linked to the Q¹ synthons through linkermolecules.

Nanofilms and Membranes from Bridged Macrocyclic Modules

In one aspect of the invention, bridged macrocyclic modules may becoupled to form a two-dimensional array or nanofilm. These nanofilms mayhave regions in which unique structures exist, which may repeat atregular intervals to provide a lattice of pores having substantiallyuniform dimensions. The unique structures may have a variety of shapesand sizes, thereby providing pores of various shapes and sizes. Becausethe unique structures may be formed in a monolayer of molecularthickness, the pores defined by the unique structures may include acavity, opening, or chamber-like structure of molecular size. Ingeneral, pores of atomic to molecular size defined by those uniquestructures may be used for selective permeation or molecular sievingfunctions. Some aspects of nanotechnology are given in NanostructuredMaterials, J. Ying, ed., Academic Press, San Diego, 2001.

Methods of Preparing Nanofilms

In one variation, bridged macrocyclic modules can be oriented on asurface or subphase by providing functional groups on the modules whichimpart amphiphilic character to the modules. For example, when themodule is deposited on a hydrophilic surface, hydrophobic substituentgroups or hydrophobic tails attached to the module may cause the moduleto reorient on the surface so that the hydrophobic substituents areoriented away from the surface, leaving a more hydrophilic facet of themodule oriented toward the surface.

The amphiphilic character may arise from, e.g., atoms in the synthons,the linkages between the synthons, functional groups coupled to thesynthons or linkages, and the bridge moiety. For example, lipophilicand/or hydrophilic moieties may be coupled to the same or differentsynthon or linkage in an amphiphilic bridged macrocyclic module.Lipophilic and hydrophilic moieties may be coupled to the macrocyclicmodule or moiety before or after formation of the closed ring of themacrocyclic module or moiety. For example, lipophilic or hydrophilicmoieties may be added to the macrocyclic module after formation of theclosed ring by substitution of a synthon or linkage, followed bycoupling of a bridge moiety to produce a bridged macrocyclic module.Examples of functional groups added to the components to impartamphiphilic character to the modules include alkyl groups, alkoxygroups, —NHR, —OC(O)R, —C(O)OR, —NHC(O)R, —C(O)NHR, —CH═CHR, and —C≡CR,where the carbon atoms of an alkyl group may be interrupted by one ormore —S—, double bond, triple bond or —SiRR— group(s), or substitutedwith one or more fluorine atoms, or any combination thereof, whereineach R is independently hydrogen or alkyl.

The amphiphilicity of a bridged macrocyclic module may be characterizedin part by its ability to form a stable Langmuir film. A Langmuir filmmay be formed on a Langmuir trough at a particular surface pressuremeasured in milliNewtons per meter (mN/m) with a particular barrierspeed measured in millimeters per minute (mm/min), and the isobariccreep or change in film area at constant surface pressure can bemeasured to characterize stability of the film. For example, a stableLangmuir film of bridged macrocyclic modules on a water subphase mayhave an isobaric creep at 5-15 mN/m such that the majority of the filmarea is retained over a period of time of about one hour. Examples ofstable Langmuir films of bridged macrocyclic modules on a water subphasemay have isobaric creep at 5-15 mN/m such that at least about 70% of thefilm area is retained over a period of time of about 30 minutes,sometimes at least about 70% of the film area is retained over a periodof time of about 40 minutes, sometimes at least about 70% of the filmarea is retained over a period of time of about 60 minutes, andsometimes at least about 70% of the film area is retained over a periodof time of about 120 minutes. In other embodiments, a stable Langmuirfilm of bridged macrocyclic modules on a water subphase may have anisobaric creep at 5-15 mN/m such that at least about 80% of the filmarea is retained over a period of time of about thirty minutes,sometimes at least about 85% of the film area is retained over a periodof time of about thirty minutes, sometimes at least about 90% of thefilm area is retained over a period of time of about thirty minutes,sometimes at least about 95% of the film area is retained over a periodof time of about thirty minutes, and sometimes at least about 98% of thefilm area is retained over a period of time of about thirty minutes.

In one example, the amphiphilic components may be dissolved in a solventand deposited on an air-subphase interface in a Langmuir trough to formthe monolayer. Typically, movable plates or barriers are used tocompress the monolayer and decrease its surface area to form a moredense monolayer. At various degrees of compression, having correspondingsurface pressures, the monolayer may reach various condensed states. Theconformation of a module on a surface may depend on the loading,density, or state of the phase or layer in which the module resides onthe surface. Surfaces which may be used to orient modules or othermolecules include interfaces such as gas-liquid, air-water, immiscibleliquid-liquid, liquid-solid, or gas-solid interfaces.

Surface pressure versus film area isotherms are obtained by the Wilhelmybalance method to monitor the state of the film. Extrapolation of theisotherm to zero surface pressure reveals the average surface area percomponent, or mean molecular area, before the components are coupled.The isotherm gives an empirical indication of the state of the thinfilm. Surface-oriented macrocyclic modules and/or other components in ananofilm layer may be in an expanded state, a liquid state, or aliquid-expanded state, or may be condensed, collapsed, or a solid phaseor close-packed state.

Oriented bridged macrocylic modules may be coupled to form a nanofilm.The modules may be oriented on the surface before or during the processof coupling. The bridged macrocyclic modules may be directly coupled toeach other, or may be coupled through linker molecules. The bridgedmacrocyclic module compounds may be coupled to one another viafunctional groups located on the synthons or on the bridge moiety.Examples of suitable functional groups and resulting linkages are shownin Tables 6-8. A bridge moiety having functional groups for couplingbridged macrocyclic module compounds may be mono-functional ormulti-functional. In one variation, a general scheme for couplingbridged macrocyclic module compounds includes coupling the functionalgroups located on the bridge moieties, as shown in Scheme 13.

wherein A indicates a bridge moiety, and X″ indicates a functional groupfor coupling complementary functional groups on other bridge moieties.

The nanofilm may be prepared from a single type of bridged macrocyclicmodule. In other variations, the nanofilm may be prepared from two ormore types of bridged macrocyclic modules.

Nanofilms may be prepared by various alternative methods. For example,linker molecules may be added to the solution containing the modules,which is subsequently deposited on the surface of the Langmuir subphase.Alternatively, the linker molecules may be added to the water subphaseof the Langmuir trough, and subsequently transfer to the layer phasecontaining the bridged macrocyclic modules for coupling. In someinstances, macrocyclic modules may be added to the subphase of theLangmuir trough, and subsequently transfer to the interface. In general,water soluable components (such as linker molecules) may be added to thesubphase for the formation of a nanofilm.

When two or more types of bridged macrocyclic modules are used informing the nanofilm, the modules may be mixed prior to or duringorientation on a surface.

Other variations will be apparent to those of skill in the art.

In general, coupling of the components of a nanofilm may be initiated bychemical, thermal, photochemical, electrochemical, and irradiativemethods. In some variations of this invention, the type of coupling ofthe components of a nanofilm may depend on the type of initiation andthe chemical process involved.

Functional groups added to the modules to impart amphiphilic charactermay in some embodiments be removed during or after formation of thenanofilm. The method of removal depends on the functional group. Thegroups attached to the modules which impart amphiphilic character to themodule may include functional groups which can be used to remove thegroups at some point during or after the process of formation of ananofilm. Acid or base hydrolysis may be used to remove groups attachedto the module via a carboxylate or amide linkage. An unsaturated grouplocated in the functional group which imparts amphiphilic character tothe module may be oxidized and cleaved by hydrolysis. Photolyticcleavage of the functional group which imparts amphiphilic character tothe module may also be done. Examples of cleavable functional groupsinclude

where n is zero to four, which is cleavable by light activation, and

where b is zero to four, and c is 7 to 27, which is cleavable by acid orbase catalyzed hydrolysis.

A variety of functional groups may be used in coupling the nanofilms.The nature of the nanofilm product formed by coupling bridgedmacrocyclic modules depends, e.g., on the specific functional groupsused, and the relative orientations of the functional groups withrespect to the module structure. The functional groups may in some casescontribute to the amphiphilic character of the module before or aftercoupling, and may be covalently or non-covalently attached to themodules. In a preferred embodiment, the functional groups are covalentlyattached to the modules. The functional groups may be attached to themodules before, during, or after orientation of the modules on a surfaceor subphase.

The coupling of modules in a nanofilm may attach two or more componentsby a linkage or linkages. The coupling may attach more than two modules,for example, by an array of linkages each formed between the modules.Each module may form one or more linkages to one or more modules, andeach module may form several types of linkages, including thoseexemplified in Tables 6-8. A module may have direct linkages, linkagesthrough a linker molecule, and linkages which include spacers, in anycombination. A linkage may connect any portion of a module to anyportion of another module. An array of linkages and an array of modulesmay be described in terms of the theory of Bravais lattices and theoriesof symmetry.

Coupling of modules may be complete or incomplete, providing a varietyof structural variations useful as nanofilm membranes. A portion of eachof the components of a nanofilm may be coupled, while the remainder ofeach is not coupled. The modules may interact through, for example,hydrogen bonding, van der Waals, and other interactions. The arrangementof linkages formed in a nanofilm may be represented by a type ofsymmetry, or may be substantially unordered.

The types of coupling between the bridged macrocyclic modules and thephase and domain behaviour of the modules, as described herein, mayinfluence the composition and properties of the product nanofilm. Amacrocyclic module may participate in more than one type of coupling.

Structure of Nanofilms

Generally, the nanofilms formed are one molecule thick throughout, butmay vary locally due to physical and chemical forces. In someembodiments, pores are supplied through the structure of the nanofilm.In some embodiments, pores are supplied through the structure of thebridged macrocyclic modules.

A wide variety of structural features and properties such as amorphous,glassy, semicrystalline or crystalline structures, and elastomeric,pliable, thermoplastic, or deformation properties may be exhibited bythe nanofilms.

The composition of the nanofilm may be solid, gel, or liquid. Themodules of the nanofilm may be in an expanded state, a liquid state, ora liquid-expanded state. The state of the modules of the nanofilm may becondensed, liquid-condensed, collapsed, or may be a solid phase orclose-packed state. The modules of the nanofilm may interact with eachother by weak forces of attraction. Alternatively, they may be coupledthrough, for example, covalent bonds. For example, the modules of ananofilm prepared from surface-oriented bridged macrocyclic modules neednot be linked by any strong interaction or coupling. Alternatively, forexample, the modules of the nanofilm may be linked through, for example,covalent bonds.

The thickness of nanofilms described herein, whether through coupled ornon-coupled bridged macrocyclic modules, is exceptionally small, oftenbeing less than about 30 nanometers, sometimes less than about 20nanometers, and sometimes from about 1-15 nanometers. The thickness of ananofilm depends partly on the structure and nature of the groups on themodules which impart amphiphilic character to the modules. The thicknessmay be dependent on temperature, and the presence of solvent on thesurface or located within the nanofilm. The thickness may be modified ifthe groups on the modules which impart amphiphilic character, inparticular the lipophilic moiety, are removed or modified after themodules have been coupled, or at other points during or after theprocess of preparation of a nanofilm. The thickness of a nanofilm mayalso depend on the structure and nature of the surface attachment groupson the components. The thickness of nanofilms may be less than about300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 or 5 Å.

Nanofilm structures define pores through which atoms, molecules, orparticles of only up to a certain size and composition may pass. Onevariation of a nanofilm structure includes an area of nanofilm able toface a fluid medium, either liquid or gaseous, and provide pores oropenings through which atoms, ions, small molecules, biomolecules, orother species are able to pass. The dimensions of the pores defined bynanofilm structures may be exemplified by quantum mechanicalcalculations and evaluations, and physical tests, as further describedin the following Examples.

The dimensions of the pores defined by nanofilm structures are describedby actual atomic and chemical structural features of the nanofilm. Theapproximate diameters of pores formed in the structure of a nanofilm arefrom about 1-150 Å, or more. In some embodiments, the dimensions of thepores are about 1-10 Å, about 3-15 Å, about 10-15 Å, about 15-20 Å,about 20-30 Å, about 30-40 Å, about 40-50 Å, about 50-75 Å, about 75-100Å, about 100-125 Å, about 125-150 Å, about 150-300 Å, about 600-1000 Å.The approximate dimensions of pores formed in the structure of ananofilm are useful to understand the porosity of the nanofilm. On theother hand, the porosity of conventional membranes is normallyquantified by empirical results such as molecular weight cut-off, whichreflects complex diffusive and other transport characteristics.

In one variation, a nanofilm structure may comprise an array of coupledmodules which provides an array of pores of substantially uniform size.The pores of uniform size may be defined by the individual modulesthemselves. Each module defines one or more pores of a particular size,depending on the conformation and state of the module. For example, theconformation of the coupled module of the nanofilm may be different fromthe nascent, pure module in a solvent, and both may be different fromthe conformation of the amphiphilic module oriented on a surface beforecoupling.

Modules of various composition and structure may be prepared whichdefine pores of different sizes. Thus, nanofilms having pores of variousdimensions are provided, depending on the particular module used toprepare the nanofilm.

In other instances, nanofilm structures define pores in the matrix ofcoupled modules. Pores defined by nanofilm structures may have a widerange of dimensions, for example, dimensions capable of selectivelyblocking the passage of small molecules or large molecules. For example,nanofilm structures may be formed from the coupling of two or moremodules, in which an interstitial pore is defined by the combinedstructure of the linked modules. A nanofilm may have an extended matrixof pores of various dimensions and characteristics. Interstitial poresmay be, for example, less than about 5 Å, less than about 10 Å, about3-15 Å, about 10-15 Å, about 15-20 Å, about 20-30 Å, about 30-40 Å,about 40-50 Å, about 50-75 Å, about 75-100 Å, about 100-125 Å, about125-150 Å, about 150-300 Å, about 300-600 Å, about 600-1000 Å.

The coupling process may result in a nanofilm in which regions of thenanofilm are not precisely monomolecular layers. Various types of localstructures are possible which do not prevent use of the nanofilm in avariety of applications. Local structural features may includeamphiphilic modules which are flipped over relative to their neighbors,or turned in a different orientation, having their hydrophobic andhydrophilic facets oriented differently than neighboring species. Localstructural features may also include overlaying or stacking of moleculesin which the nanofilm is two or more molecular layers thick, localregions in which the interlinking of the modules is not complete so thatsome of the available coupling groups are not coupled to other species,or local regions in which there is an absence of a particular molecule.Other local structural features may include grain boundaries andorientational faults. In one variation, the nanofilm has a thickness ofup to 30 nanometers due to the layering of nanofilm structures.

The nanofilms disclosed herein may be substantially uniform with respectto the orientation of their amphiphilic modules, but may in someembodiments comprise regions of local structural features as indicatedhereinabove. Local structural features may comprise, for example,greater than about 30%, less than about 30%, less than about 20%, lessthan about 15%, less than about 10%, less than about 5%, less than about3%, less than about 1% of the surface area of the nanofilm.

A nanofilm may have an array of coupled modules in which the positionalordering of the modules is random, or is non-random with regions inwhich one type of species is predominant.

In alternative variations, the nanofilm may include additives,dispersants, surfactants, excipients, compatiblizers, emulsifiers,suspension agents, plasticizers, or other species which modify theproperties of the nanofilm. In some instances, the nanofilm may bederivatized to provide biocompatability or reduce fouling of thenanofilm by attachment or adsorption of biomolecules.

Substrates

Nanofilms may be deposited on a substrate by various methods, such asLangmuir-Schaefer, Langmuir-Blodgett, or other methods used withLangmuir systems. In one variation, a nanofilm is deposited on asubstrate in a Langmuir tank by locating the substrate in the subphasebeneath the air-water interface, and lowering the level of the subphaseuntil the nanofilm lands gently on the substrate and is thereforedeposited. A description of Langmuir films and substrates is given inU.S. Pat. Nos. 6,036,778, 4,722,856, 4,554,076, and 5,102,798, and in R.A. Hendel et al., Vol. 119, J. Am. Chem. Soc. 6909-18 (1997). Adescription of films on substrates is given in Munir Cheryan,Ultrafiltration and Microfiltration Handbook (1998).

Other methods for preparing nanofilms include forced removal of solventto prepare a film, such as spin coating methods and spray coatingmethods, as well as coating and deposition methods includinginterfacial, dip coating, knife-edge coating, grafting, casting, phaseinversion, or electroplating or other plating methods.

Nanofilms deposited on a substrate may be cured or annealed by chemical,thermal, photochemical, electrochemical, irradiative or drying methodsduring or after deposition on a substrate. For example, chemical methodsinclude reactions with vapor phase reagents such as ethylenediamine orsolution phase reagents. A nanofilm treated by any method to attach orcouple it to a substrate may be said to be cured.

The deposition may result in non-covalent or weak attachment of thenanofilm to the substrate through physical interactions and weakchemical forces such as van der Waals forces and weak hydrogen bonding.The nanofilm may in some embodiments be bound to the substrate throughionic or covalent interaction, or other type of interaction.

The substrate may be any surface of any material. Substrates may beporous or non-porous, and may be made from polymeric and inorganicsubstances. Examples of porous substrates include plastics or polymers,track-etch polycarbonate, track-etch polyester, polyethersulfone,polysulfone, gels, hydrogels, cellulose acetate, polyamide, PVDF,polyethylene terephthalate or polybutylene terephthalate, polyvinylchloride, polyvinylidene chloride, polytetrafluoroethylene, polyethyleneor polypropylene, ceramics, anodic alumina, laser ablated and otherporous polyimides, and UV etched polyacrylate. Examples of non-poroussubstrates include silicon, germanium, glass, metals such as platinum,nickel, palladium, aluminum, chromium, niobium, tantalum, titanium,steel, or gold, glass, silicates, aluminosilicates, non-porous polymers,and mica. Further examples of substrates include diamond and indium tinoxide. Preferred substrates include silicon, gold, SiO₂,polyethersulfone, and track etch polycarbonate. In some embodiments, thesubstrate is SiO₂. In other embodiments, the substrate is polycarbonatetrack etch membrane.

Substrates may have any physical shape or form including films, sheets,plates, or cylinders, and may be particles of any shape or size.

A nanofilm deposited on a substrate may serve as a membrane. Any numberof layers of nanofilm may be deposited on the substrate to form amembrane. In some variations, a nanofilm is deposited on both sides of asubstrate.

A layer or layers of various spacing materials may be deposited orattached in between layers of a nanofilm, and a spacing layer may alsobe used in between the substrate and the first deposited layer ofnanofilm. Examples of spacing layer compositions include polymericcompositions, hydrogels (acrylates, poly vinyl alcohols, polyurethanes,silicones), thermoplastic polymers (polyolefins, polyacetals,polycarbonates, polyesters, cellulose esters), polymeric foams,thermosetting polymers, hyperbranched polymers, biodegradable polymerssuch as polylactides, liquid crystalline polymers, polymers made by atomtransfer radical polymerization (ATRP), polymers made by ring openingmetathesis polymerization (ROMP), polyisobutylenes and polyisobutylenestar polymers, and amphiphilic polymers. Other examples of spacing layercompositions include inorganics, such as inorganic particles such asinorganic microspheres, colloidal inorganics, inorganic minerals, silicaspheres or particles, silica sols or gels, clays or clay particles, andthe like. Examples of amphiphilic molecules include amphiphilescontaining polymerizable groups such as diynes, enes, or amino-esters.The spacing layers may serve to modify barrier properties of thenanofilm, or may serve to modify transport, flux, or flowcharacteristics of the membrane or nanofilm. Spacing layers may serve tomodify functional characteristics of the membrane or nanofilm, such asstrength, modulus, or other properties.

In some variations, a nanofilm may be deposited on a surface and adhereto the surface to a degree sufficient for many applications, such asfiltration and membrane separations, without coupling to the surface.

In other variations, a nanofilm may be, for example, covalently coupledto a substrate surface. Surface attachment groups may be provided on thebridge moiety or synthons of a bridged macrocyclic module, which may beused to couple the nanofilm to the substrate.

Examples of functional groups which may be used as surface attachmentgroups to couple a nanofilm to a substrate include amine groups,carboxylic acid groups, carboxylic ester groups, alcohol groups, glycolgroups, vinyl groups, styrene groups, olefin styryl groups, epoxidegroups, thiol groups, magnesium halo or Grignard groups, acrylategroups, acrylamide groups, diene groups, aldehyde groups, and mixturesthereof.

A substrate may have functional groups which couple to the functionalgroups of a nanofilm. The functional groups of the substrate may besurface groups or linking groups bound to the substrate, which may beformed by reactions which bind the surface groups or linking groups tothe substrate. Surface groups may also be created on the substrate by avariety of treatments such as cold plasma treatment, surface etchingmethods, solid abrasion methods, or chemical treatments. Some methods ofplasma treatment are given in Inagaki, Plasma Surface Modification andPlasma Polymerization, Technomic, Lancaster, Pa., 1996. In someembodiments, the substrate is derivatized with APTES. In otherembodiments, the substrate is derivatized withmethylacryloxymethyltrimethoxysilane (MAOMTMOS). In other embodiments,the substrate is derivatized with acryloxypropyltrimethoxy-silane(AOPTMOS).

Surface attachment groups of the nanofilm and the surface may be blockedwith protecting groups until needed. Non-limiting examples of suitablefunctional groups for coupling the nanofilm to the substrate and theresulting linkages may be found in Tables 6-8.

Surface attachment groups may be connected to a nanofilm by spacergroups. Likewise, substrate functional groups may be connected to thesubstrate by spacer groups. Spacer groups for surface attachment groupsmay be polymeric. Examples of polymeric spacers include polyethyleneoxides, polypropylene oxides, polysaccharides, polylysines,polypeptides, poly(amino acids), polyvinylpyrrolidones, polyesters,polyvinylchlorides, polyvinylidene fluorides, polyvinylalcohols,polyurethanes, polyamides, polyimides, polysulfones, polyethersulfones,polysulfonamides, and polysulfoxides. Examples of polymeric spacerstructures include linear, branched, comb and dendrimeric polymers,random and block copolymers, homo- and heteropolymers, flexible andrigid chains. Spacer groups for surface attachment groups may alsoinclude bifunctional linker groups or heterobifunctional linker groupsused to couple biomolecules and other chemical species.

In one variation, a photoreactive group such as a benzophenone or otherlight activated cross-linker is bound to the substrate. Thephotoreactive group may be activated with light, for example,ultraviolet light, to provide a reactive species which couples to ananofilm.

Surface attachment of modules may also be achieved throughligand-receptor mediated interactions, such as biotin-streptavidin. Forexample, the substrate may be coated with streptavidin, and biotin maybe attached to the modules, for example, through linker groups such asPEG or alkyl groups.

Membranes and Filtration Function

The nanofilms described herein may be useful, for example, as membranes.The membrane may be brought into contact with a fluid or solution,separating a species or component from that fluid or solution, forexample, for purposes of filtration. Normally, a membrane is a substancewhich acts as a barrier to block the passage of some species, whileallowing restricted or regulated passage of other species. In general,permeants may traverse the membrane if they are smaller than a cut-offsize, or have a molecular weight smaller than a so-called cut-offmolecular weight. The membrane may be called impermeable to specieswhich are larger than the cut-off molecular weight. The cut-off size ormolecular weight is a characteristic property of the membrane. Selectivepermeation is the ability of the membrane to cut-off, restrict, orregulate passage of some species, while allowing smaller species topass. Thus, the selective permeation of a membrane may be describedfunctionally in terms of the largest species able to pass the membraneunder given conditions. The size or molecular weight of various speciesmay also be dependent on the conditions in the fluid to be separated,which may determine the form of the species. For example, species mayhave a sphere of hydration or solvation in a fluid, and the size of thespecies in relation to membrane applications may or may not include thewater of hydration or the solvent molecules. Thus, a membrane ispermeable to a species of a fluid if the species can traverse themembrane in the form in which it normally would be found in the fluid.Permeation and permeability may be affected by interaction between thespecies of a fluid and the membrane itself. While various theories maydescribe these interactions, the empirical measurement of pass/no-passinformation relating to a nanofilm, membrane, or module is a useful toolto describe permeation properties. A membrane is impermeable to aspecies if the species cannot pass through the membrane.

Pores may be provided in the nanofilms described herein, for example,pores may be supplied in the structure of the nanofilm. Pores may besupplied in the structure of the bridged macrocyclic modules. The typeand degree of crosslinking between modules may influence pore size.

The nanofilms may have molecular weight species cut offs of, forexample, greater than about 15 kDa, greater than about 10 kDa, greaterthan about 5 kDa, greater that about 1 kDa, greater than about 800 Da,greater than about 600 Da, greater than about 400 Da, greater than about200 Da, greater than about 100 Da, greater than about 50 Da, greaterthan about 20 Da, less than about 15 kDa, less than about 10 kDa, lessthan about 5 kDa, less that about 1 kDa, less than about 800 Da, lessthan about 600 Da, less than about 400 Da, less than about 200 Da, lessthan about 100 Da, less than about 50 Da, less than about 20 Da, about13 kDa, about 190 Da, about 100 Da, about 45 Da, about 20 Da.

“High permeability” indicates a clearance of, for example, greater thanabout 70%, greater than about 80%, or greater than about 90% of thesolute. “Medium permeability” indicates a clearance of, for example,less than about 50%, less than about 60%, or less than about 70% of thesolute. “Low permeability” indicates a clearance of less than, forexample, about 10%, less than about 20%, or less than about 30% of thesolute. A membrane is impermeable to a species if it has a very lowclearance (for example, less than about 5%, less than about 3%) for thespecies, or if it has very high rejection for the species (for example,greater than about 95%, greater than about 98%). The passage orexclusion of a solute is measured by its clearance, which reflects theportion of solute that actually passes through the membrane. Forexample, the no pass symbol in Tables 12-13 indicates that the solute ispartly excluded by the module, sometimes less than about 90% rejection,often at least about 90% rejection, sometimes at least about 98%rejection. The pass symbol indicates that the solute is partly clearedby the module, sometimes less than about 90% clearance, often at leastabout 90% clearance, sometimes at least about 98% clearance.

Examples of processes in which nanofilms may be useful include processesinvolving liquid or gas as a continuous fluid phase, filtration,clarification, fractionation, pervaporation, reverse osmosis, dialysis,hemodialysis, affinity separation, oxygenation, and other processes.Filtration applications may include ion separation, desalinization, gasseparation, small molecule separation, separation of enantiomers,ultrafiltration, microfiltration, hyperfiltration, water purification,sewage treatment, removal of toxins, removal of biological species suchas bacteria, viruses, or fungus.

Networked Arrays from Bridged Macrocyclic Module Compounds

In another aspect of the invention, bridged macrocyclic modules may becoupled to form a networked array, such as a 3-D array. The 3-D arraysmay also be useful as nanofilms.

In one variation, a general scheme for coupling bridged macrocyclicmodule compounds in a network includes coupling the functional groupslocated on the bridge moieties, as shown in Scheme 14.

In Scheme 14, A is a bridge moiety, and X″ are independently selectedfunctional groups for coupling to corresponding functional groups onother bridged macrocyclic modules. Suitable examples of functionalgroups, linkers, and the resulting linkages may be found in Tables 6-8.

Networked arrays may be made, for example, through polymerizationschemes, such as those shown in Schemes 1-3.

The following examples further describe and demonstrate variationswithin the scope of the present invention. All examples described inthis specification, both in the description above and the examplesbelow, are given solely for the purpose of illustration and are not tobe construed as limiting the present invention. While there have beendescribed illustrative variations of this invention, those skilled inthe art will recognize that they may be changed or modified withoutdeparting from the spirit and scope of this invention, and it isintended to cover all such changes, modifications, and equivalentarrangements that fall within the true scope of the invention as setforth in the appended claims.

All documents referenced herein, including applications for patent,patent references, publications, articles, books, and treatises, arespecifically incorporated by reference herein in their entirety.

EXAMPLES

Reagents were obtained from Aldrich Chemical Company (St. Louis, Mo.).The Langmuir trough used was a KSV minitrough (KSV Instruments,Trumbull, Conn.). Rates of surface compression are reported as thelinear rate of barrier movement. Atomic force microscopy (AFM) imagesmay be obtained with a PicoSPM (Molecular Imaging, Pheonix Ariz.).Contact Mode images may be recorded under flowing nitrogen with an Sipoint probe tip.

Example 1

Bridged macrocyclic module compound 6 was prepared as shown in Scheme15.

Diprotected 4-methyldialdehyde phenol (1). To a 250 mL Schlenk flaskwith stirbar under argon 2-hydroxy-5-methyl-1,3-benzenedicarboxaldehyde(15.2 mmol, 2.5 g) was added and the flask evacuated and backfilled withargon 3×. Anhydrous benzene (80 mL) was cannula transferred followed bystirring at ambient temperature. Next, anhydrous ethylene glycol (91.4mmol, 5.67 g) was added via syringe under argon followed by TsOH (0.167mmol, 0.032 g). The reaction vessel was fitted with a Dean-Stark trapand reflux condenser and the reaction refluxed for ca. 23 h. Thereaction was diluted with ethyl acetate (300 mL) washed with 1M NaHCO₃(50 mL), H₂O (50 mL), then brine (50 mL). The organic layer wasseparated and dried over Na₂SO₄, filtered and the solvent removed byrotovaporation. Purification by silica gel chromatography (2:1hexane:ethyl acetate) afforded a white solid (2.34 g; 61% yield). IR(cm⁻¹) 3329, 2917, 2895, 1627, 1496, 1478, 1408, 928, 873; ¹H NMR (400MHz, CDCl₃) δ 8.07 (s, 1H, OH), 7.18 (s, 2H, ArH), 6.03 (s, 2H, ArCHO₂),4.20-4.00 (m, 8H, OCH₂CH₂O), 2.26 (s, 3H, ArMe); {¹H} ¹³C NMR (100 MHz,CDCl₃) 151.8, 129.1, 122.0, 102.1, 65.3, 20.8.

Triethylene glycol tethered diprotected2-hydroxy-5-methyl-1,3-benzenedicarboxaldehyde (2). To a 25 mL schlenkflask with Kontes valve and stirbar under argon 1 (1.67 mmol, 0.423 g)was added and the flask evacuated and backfilled with argon 3×.Anhydrous DMF (12.2 mL) was added via syringe and the solution stirredat rt. Next, anhydrous CsCO₃ (3.52 mmol, 1.15 g) was added followed bytriethylene glycol ditosylate (0.808 mmol, 0.371 g) and the mixturestirred at rt. The vessel was closed, stirred and heated to 70° C. forca. 12 h. The reaction was allowed to cool to rt then diluted with ethylacetate (10 mL), filtered and the solid washed with ethyl acetate (3×100mL) and filtered. The organic extracts were combined and washed withsaturated NH₄Cl(aq) (25 mL) then washed with brine (3×25 mL) dried overNa₂SO₄, filtered and the solvent removed by rotovaporation to afford thecrude product. Purification by silica plug (ethyl acetate) afforded awhite solid (0.464 g; 89% yield). IR (cm⁻¹) 2947, 2884, 1683, 1601,1476, 1397, 1110; ¹H NMR (400 MHz, CDCl₃) δ 7.38 (s, 4H, ArH), 6.13 (s,4H, ArCHO₂), 4.18 (m, 4H, ArOCH₂—), 4.02 (m, 16H, OCH₂CH₂O), 3.84 (m,4H, ArOCH₂CH₂—), 3.78 (s, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 2.33 (s, 6H,ArCH₃); {¹H} ¹³C NMR (100 MHz, CDCl₃) 154.5, 134.2, 130.8, 129.0, 99.1,76.0, 70.9, 70.5, 65.5, 21.2.

Triethylene glycol tethered2-hydroxy-5-methyl-1,3-benzenedicarboxaldehyde (3). To a 1000 mLpear-shaped flask and 100 mL pear shaped flask and stirbar under argon 2(3.20 mmol, 1.98 g and 0.488 mmol, 0.302 g, respectively), were added.The substrate was dissolved in THF (160 mL, 24.4 mL, respectively) then5% HCl(aq) (56.8 mL, 8.7 mL) was added and the mixture stirred for 12 hresulting in a white precipitate. The reactions were combined and pouredinto a separatory funnel containing 50 mL of NaHCO₃ (sat, aq) thereaction vessel rinsed with 100 mL of ethyl acetate poured into theseparatory funnel with an additional 250 mL of ethyl acetate and theaqueous layer extracted, separated and washed with brine (3×50 mL). Theorganic layer was dried over Na₂SO₄, filtered and the filter and dryingagent washed with CHCl₃, the solvents combined and volatiles removed byrotovaporation. Purification by silica gel plug (dissolve in CHCl₃,ethyl acetate as mobile phase) afforded a white solid (1.63 g, 99%). IR(cm⁻¹) 3347, 2947, 2911, 2856, 2760, 1678, 1580, 1465, 1404, 1130; ¹HNMR (400 MHz, CDCl₃) δ 10.43 (s, 4H, ArCHO), 7.87 (s, 4H, ArH), 4.31 (m,4H, ArOCH₂—), 3.67 (s, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 2.40 (s, 6H,ArCH₃); {¹H} ¹³C NMR (100 MHz, CDCl₃) 189.6, 162.5, 135.4, 135.0, 130.1,78.6, 71.0, 70.4, 20.8.

Triethylene glycol tethered 2-hydroxy-5-methyl-1,3-benzenedicarboxylicacid (4). To a 100 mL round bottom flask and stirbar under argon 3 (2.43mmol, 1.08 g) was added. The substrate was dissolved in 1,4-dioxane (186mL) then sonicated until homogeneous. Buffer, NaH₂PO₄ (37.9 mmol, 4.55g) and sulfamic acid (14.6 mmol, 1.42 g) were dissolved in de-ionizedH₂O in a 41.1 mL Erlenmeyer flask with a stirbar, then transferred tothe stirring solution of the substrate by syringe. The heterogeneoussolution was sonicated again until the mixture was nearly homogeneous.Sodium chlorite (NaClO₂, 12.6 mmol, 1.14 g) was dissolved in ade-ionized H₂O (12.6 mL) and added dropwise via syringe to the stirringsolution resulting in a yellow homogeneous solution. After ca. 3.5 h thereaction was complete as assessed by TLC. Sodium sulfite (11.7 mmol,1.47 g) was added, and the reaction stirred for ca. 1 h giving acolorless solution. The reaction mixture was diluted with CH₂Cl₂ (500mL), poured into a separatory funnel, H₂O (200 mL) was added, and the pHadjusted to ca. 1-2 with 4 M HCl. The organic layer was separated andwashed with brine (3×50 mL). The organic layer was again separated, andthe solvent removed by rotovaporation and put under vacuo to remove theremaining H₂O affording a white solid (1.22 g, 99% yield). IR (cm⁻¹)3060, 2960, 2923, 2853, 2687, 2568, 1678, 1604, 1578, 1467, 1450, 1257,1244, 1118, 1102; ¹H NMR (400 MHz, DMSO-d₆) δ 13.04 (s, 4H, ArCOOH),7.60 (s, 4H, ArH), 4.06 (m, 4H, ArOCH₂—), 4.06 (m, 4H, ArOCH₂CH₂—), 3.54(s, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 2.30 (s, 6H, ArCH₃); {¹H} ¹³C NMR(100 MHz, DMSO-d₆) 167.1, 154.4, 133.7, 132.9, 127.5, 74.3, 69.6, 69.4,66.4, 19.9.

Triethylene glycol tethered 2-hydroxy-5-methyl-1,3-benzenediamidetetra-cis-1,3-diaminocyclohexane (5). To a 50 mL round bottom Schlenkflask with Kontes valve and stirbar under argon 4 (0.096 mmol, 0.100 g)was added. The vessel was evacuated and backfilled with argon (3×).Oxalyl chloride (2.0 M) in dichloromethane (0.985 mmol, 0.480 mL) wasadded via syringe. Next a catalytic amount of DMF (0.015 mL) was addedvia syringe with stirring at rt for 2 h, resulting in rigorouseffervescence and a yellow solution with yellow precipitate. Allvolatiles were removed under vacuo and the residue dissolved indichloromethane (1 mL) followed by addition of triethylamine (1.53 mmol,0.155 g) via syringe under argon. A dichloromethane solution of mono Bocprotected cis-1,3-diaminocyclohexane (0.980 mmol, 0.210 g) was addeddropwise to the reaction mixture, the vessel sealed and stirredovernight at ambient temperature affording an orange solution withprecipitate formation. The reaction was diluted with dichloromethane(250 mL), washed with brine (2×50 mL), water (50 mL), then brine (50mL), the organic layer dried over sodium sulfate, filtered and thesolvent removed by rotovaporation. Purification by silica chromatography(15:1:1 ethyl acetate:hexane) afforded an off white solid (0.216, 89%yield). IR (cm⁻¹) 3307, 3060, 2967, 2934, 2853, 1682, 1639, 1600, 1520,1450, 868; ¹H NMR (400 MHz, CDCl₃) δ 7.70 (bs, 4H, ArH), 7.25 (bs, 4H,ArH) 4.73 (bs, 4H, ArC(O)NH), 4.05 (m, 4H, ArOCH₂CH₂—), 3.99 (bs, 4H,CHNHC(O)O^(t)—Bu), 3.69 (m, 4H, ArOCH₂CH₂—), 3.60 (bs, 4H,ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 3.53 (bs, 4H, cyclohexane), 2.30 (m, 4H,cyclohexane), 2.02 (m, 7H, cyclohexane), 1.83, 1.80 (m, s 11H,cyclohexane), 1.43 (bs, 36H NHC(O)OC(CH₃)₃), 1.20-1.00 (m, 10H,cyclohexane); ESIMS (MNa+)=1314 m/z.

Bridged macrocyclic module (6): To a 15 mL round bottom flask withstirbar under argon 5 (0.0107 mmol, 0.0138 g) was added. The vessel wasevacuated and backfilled with argon (3×). The substrate was dissolved indichloromethane (0.712 mL, 0.015 M). Anhydrous trifluoroacetic acid(0.356 mL) was added via syringe, the vessel sealed under argon with aglass stopper and stirred at rt for ca. 1 h. All volatiles were removedunder vacuo followed by addition of ca. 1 mL of diethyl ether. Next,anhydrous NEt₃ (0.020 mL) was added via syringe under argon at rt andthe mixture stirred for ca. 2 h. The resulting precipitate was washedwith anhydrous diethyl ether (4×2 mL), followed by cannula filtrationand drying in vacuo to afford a light yellow solid. In a nitrogenatmosphere glovebag, the solid was dissolved in CD₃OD (2 mL) andtransferred to a vial containing2-hydroxy-5-methyl-1,3-benzenedicarboxaldehyde (0.0214 mmol, 0.0035 g).The reaction mixture was transferred to a J-Young tube, sealed andheated to 66° C. for 22 h. Purification by silica plug (4:2:0.3chloroform:hexane:triethylamine) afforded a yellow-green solid (0.0103g, 84% yield). NMR (400 MHz, CDCl₃) δ 8.70 (bs, 1H, —N═CH—), 8.69 (bs,1H, —N═CH—), 7.70 (bs, 1H, —N═CH—), 7.69 (bs, 1H, —N═CH—), 7.65-7.35 (m,4H, ArH), 7.11 (bs, 4H, 5.54 (bs, 4H, CHNHC(O)Ar), 4.07 (m, 4H,ArOCH₂CH₂—), 3.93 (m, 4H, CHNHC(O)O^(t)-Bu), 3.80-3.69 (m, 4H,ArOCH₂CH₂—), 3.62 (bs, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 2.50 (bs, 4H,cyclohexane), 2.34, 2.33, 2.28, 2.23 (bs, 12H, ArMe), 2.00 (m, 7H,cyclohexane), 1.89 (m, s 11H, cyclohexane), 1.20-1.00 (m, 10H,cyclohexane); ESIMS (MH+)=1148.9 m/z.

Example 2

Bridged macrocyclic module compound 12 was made according to Scheme 16.

In Scheme 16, R is —CH₂CO₂(CH₂)₁₅CH₃.

Diprotected Dialdehyde j (7). To a 250 mL Schlenk flask with stirbarunder argon dialdehyde j (2,6-diformyl-4-hexadecyl benzylphenolcarboxylate) (2.31 mmol, 1.0 g) was added, and the flask evacuated andbackfilled with argon 3×. Anhydrous benzene (100 mL) was cannulatransferred, followed by stirring at ambient temperature. Next,anhydrous ethylene glycol (30.48 mmol, 1.9 g) was added via syringeunder argon followed by TsOH (0.089 mmol, 0.017 g). The reaction vesselwas fitted with a Dean-Stark trap and reflux condenser and the reactionrefluxed for ca. 23 h. The reaction was complete as assessed by TLC(Rf=0.63, 1:1 hexane:ethyl acetate, KMnO₄ stain). Purification by silicagel chromatography (2:1 hexane:ethyl acetate) afforded a white solid(0.770 g; 64% yield). IR (cm⁻¹) 3389, 2917, 2850, 1729, 1627, 1615 ¹HNMR (400 MHz, CDCl₃) δ 8.19 (s, 1H, OH), 7.32 (s, 2H, ArH), 6.06 (s, 2H,ArCHO₂), 4.20-4.00 (m, 8H, OCH₂CH₂O), 3.54 (s, 2H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 1.61 (t, 2H, ³J=6.6 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃),1.27 (m, 28H, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 0.89 (t, 3H, ³J=6.6 HzCH₂C(O)OCH₂(CH₂)₁₄CH₃); {¹H} ¹³C NMR (100 MHz, CDCl₃) 171.9, 153.1,129.5, 125.5, 123.5, 101.9, 65.2, 40.7, 32.1, 29.92, 29.89, 29.88,29.81, 29.7, 29.6, 29.5, 28.8.

Triethylene glycol tethered Diprotected Dialdehyde j (8). To a 25 mLSchlenk flask with Kontes valve and stirbar under argon 7 (2.04 mmol,1.06 g) was added and the flask evacuated and backfilled with argon 3×.Anhydrous DMF (14.9 mL) was added via syringe and the solution stirredat rt. Next, anhydrous CsCO₃ (4.28 mmol, 1.39 g) was added, followed bytriethylene glycol ditosylate (0.983 mmol, 0.451 g) and the mixturestirred at rt. The vessel was closed, stirred and heated to 70° C. forca. 12 h. The reaction was allowed to cool to rt then diluted with ethylacetate (10 mL), filtered and the solid washed with ethyl acetate (3×100mL) and filtered. The organic extracts were combined and washed withsaturated NH₄Cl(aq) (75 mL), then washed with brine (3×75 mL), driedover Na₂SO₄, filtered and rotovapped to afford the crude product.Purification by column chromatography (2:1 ethyl acetate:hexane)afforded a white solid (1.035 g; 91% yield). IR (cm⁻¹) 2954, 2916, 2849,1739, 1602; ¹H NMR (400 MHz, CDCl₃) δ 7.50 (s, 4H, ArH), 6.15 (s, 4H,ArCHO₂), 4.20 (t, 4H, ³J=4.8 Hz, ArOCH₂—), 4.15-3.70 (m, 16H, OCH₂CH₂O),3.84 (t, 4H, ³J=4.8 Hz, ArOCH₂CH₂—), 3.78 (s, 4H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.59 (s, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 1.60(t, 4H, ³J=6.8 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃), 1.26 (m, 56H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 0.88 (t, 6H, ³J=6.6 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃);{¹H} ¹³C NMR (100 MHz, CDCl₃) 171.5, 155.7, 131.5, 130.3, 129.5, 99.1,76.1, 71.0, 70.5, 65.5, 65.3, 41.0, 32.1, 29.89(br), 29.86, 29.80, 29.7,29.6, 29.5, 28.7.

Triethylene glycol tethered Dialdehyde j (9). To a 200 mL pear-shapedflask and stirbar under argon 8 (0.89 mmol, 1.03 g) was added. Thesubstrate was dissolved in THF (79.4 mL), then 5% HCl (28.4 mL, aq) wasadded and the mixture stirred for 12 h resulting in a white precipitate.The reaction mixture was poured into a separatory funnel containing 50mL of NaHCO₃ (sat, aq) the reaction vessel rinsed with 100 mL of ethylacetate and poured into the separatory funnel with an additional 250 mLof ethyl acetate, and the aqueous layer extracted, separated and washedwith brine (3×50 mL). The organic layer was dried over Na₂SO₄, filteredand the solvent removed by rotovaporation. Purification by silica gelcolumn chromatography (3:1 hexane:ethyl acetate) afforded a white solid(0.864 g, 99%). IR (cm⁻¹) 2956, 2917, 2851, 1720, 1694, 1683; ¹H NMR(400 MHz, CDCl₃) δ 10.4 (s, 4H, ArCHO), 7.98 (s, 4H, ArH), 4.33 (t, 4H,³J=4.4 Hz, ArOCH₂CH₂—), 4.09 (t, 4H, ³J=6.8 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃),3.84 (t, 4H, ³J=4.4 Hz, ArOCH₂CH₂—), 3.67 (s, 4H,ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 3.65 (s, 4H, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 1.60(dt, 4H, ³J=6.8 Hz CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 1.25 (m, 52H,CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 0.88 (t, 6H, ³J=6.6 HzCH₂C(O)OCH₂(CH₂)₁₄CH₃); {¹H} ¹³C NMR (100 MHz, CDCl₃) 189.2, 170.8,163.4, 135.9, 131.4, 130.4, 78.6, 70.9, 70.4, 65.7, 65.3, 40.2, 32.1,29.89(br), 29.88, 29.86, 29.8, 29.7, 29.6, 29.4, 28.7.

Triethylene glycol tethered Dicarboxylic acid j (10). To a 100 mL roundbottom flask and stirbar under argon 9 (0.204 mmol, 0.200 g) was added.The substrate was dissolved in 1,4-dioxane (21 mL), then sonicated untilhomogeneous. Buffer, NaH₂PO₄ (3.19 mmol, 382 mg) and sulfamic acid (1.23mmol, 0.119 mg) were dissolved in de-ionized H₂O in a 25 mL Erlenmeyerflask with a ‘flea’ stirbar, then transferred to the stirring solutionof the substrate by syringe. The heterogeneous solution was sonicatedagain until the mixture was nearly homogeneous. Sodium chlorite (NaClO₂,1.06 mmol, 0.96 mg) was dissolved in a de-ionized H₂O (1 mL) and addeddropwise via syringe to the stirring solution resulting in a yellowhomogeneous solution. After ca. 3.5 h the reaction was complete asassessed by TLC. Sodium sulfite (0.98 mmol, 124 mg) was added and thereaction stirred for ca. 1 h giving a colorless solution. The reactionmixture was diluted with CH₂Cl₂ (200 mL), poured into a separatoryfunnel and the pH adjusted to ca. 1-2 with 4 M HCl. The aqueous layerwas extracted with CH₂Cl₂ (2×50 mL). The organic layer was separated andwashed with brine (3×50 mL). The organic layer was separated and thesolvent removed by rotovaporation and then put under vacuo to remove theremaining H₂O, affording a white solid (0.211 g, 99% yield). IR (cm⁻¹)3193, 2953, 2917, 2850, 1734 (br), 1607, 1582, 1467; ¹H NMR (400 MHz,DMSO-d₆) δ 13.18 (bs, 4H, ArCOOH), 7.70 (s, 4H, ArH), 4.08 (t, 4H,³J=5.1 Hz, ArOCH₂CH₂—), 4.04 (t, 4H, ³J=6.8 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃),3.69 (t, 4H, ³J=5.1 Hz, ArOCH₂CH₂—), 3.57 (s, 4H,ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 3.54 (s, 4H, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 1.54(tt, 4H, ³J=6.8 Hz CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 1.22 (m, 52H,CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 0.84 (t, 6H, ³J=6.6 HzCH₂C(O)OCH₂(CH₂)₁₄CH₃); {¹H} ¹³C NMR (100 MHz, DMSO-d₆) 171, 166.9,155.6, 134.4, 129.8, 127.6, 74.4, 69.6, 69.5, 66.4, 64.4, 31.3, 29.08(br), 29.05, 29.0, 28.9, 28.8, 28.7, 28.1, 25.3, 22.1, 14.0.

Triethylene glycol tethered Diamidetetra-Boc-protected-cis-1,3-diaminocyclohexane j (11). To a 50 mL roundbottom Schlenk flask with Kontes valve and stirbar under argon 10 (0.096mmol, 0.100 g) was added. The vessel was evacuated and backfilled withargon (3×). Oxalyl chloride (2.0 M) in dichloromethane (0.985 mmol,0.480 mL) was added via syringe. Next a catalytic amount of DMF (0.015mL) was added via syringe with stirring at rt for 2 h, resulting inrigorous effervescence and a yellow solution with yellow precipitate.All volatiles were removed under vacuo and the residue dissolved indichloromethane (1 mL) followed by addition of triethylamine (1.53 mmol,0.155 g) via syringe under argon. A dichloromethane solution ofmono-Boc-protected-cis-1,3-diaminocyclohexane (0.980 mmol, 0.210 g) wasadded dropwise to the reaction mixture, the vessel sealed and stirredovernight at ambient temperature, affording an orange solution withprecipitate formation. The reaction was diluted with dichloromethane(250 mL), washed with brine (2×50 mL), water (50 then brine (50 mL), theorganic layer dried over sodium sulfate, filtered and the solventremoved by rotovaporation. Purification by silica chromatography (2.5:1ethyl acetate:hexane) afforded an off white solid (0.154 mg, 88% yield).IR (cm⁻¹) 3360, 3320, 3053, 2924, 2854, 1710, 1696, 1642, 1520, 903,874; ¹H NMR (400 MHz, CDCl₃) δ 7.80 (bs, 4H, ArH), 7.21 (bs, 4H,ArC(O)NH), 4.75 (bs, 4H, CHNHC(O)O^(t)-Bu), 4.06 (m, 4H, ArOCH₂CH₂—),3.99 (m, 4H, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.70 (m, 4H, ArOCH₂CH₂—), 3.59 (bs,4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 3.54 (bs, 4H, CH₂C(O)OCH₂(CH₂)₁₄CH₃),2.31 (m, 4H, cyclohexane), 2.02 (m, 7H, cyclohexane), 1.81, 1.71 (m, s11H, cyclohexane), 1.61 (m, 4H CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 1.26 (m, 52H,CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 1.20-1.00 (m, 10H, cyclohexane), 0.88 (m, 6H,³J=6.6 Hz CH₂C(O)OCH₂(CH₂)₁₄CH₃); {¹H} ¹³C NMR (100 MHz, CDCl₃) 171.3,164.6, 155.3, 153.3, 134.6, 130.9, 129.0, 70.3, 70.0, 65.6, 48.8, 48.2,40.2, 39.8, 32.9, 32.2, 32.1, 29.92(br), 29.88, 29.83, 29.7, 29.6, 29.5,28.7, 28.6, 26.1 23.2, 22.9, 14.4; ESIMS (MH+)=1829.5 m/z.

Octamer IV pjs (12). To a 50 mL round bottom Schlenk flask with Kontesvalve and stirbar under argon 11 (0.0107 mmol, 0.0195 g) was added. Thevessel was evacuated and backfilled with argon (3×). The substrate wasdissolved in dichloromethane (0.712 mL. 0.015 M). Anhydroustrifluoroacetic acid (0.356 mL) was added via syringe, the vessel sealedunder argon with a glass stopper, and stirred at it for ca. 1 h. Allvolatiles were removed under vacuo, followed by addition of about 1 mLof diethyl ether. Next, anhydrous NEt₃ (0.020 mL) was added via syringeunder argon at rt, and the mixture stirred for ca. 2 h. The resultingprecipitate was washed with anhydrous diethyl ether (3×2 mL), followedby cannula filtration and drying in vacuo to afford a light yellowsolid. In a nitrogen atmosphere glovebag, the solid was dissolved inCD₃OD (2 mL) and transferred to a vial containing dialdehyde j(2,6-diformyl-4-hexadecyl benzylphenol carboxylate) (0.0214 mmol, 0.0092g). The reaction mixture was transferred to a J-Young tube, sealed andheated to 66° C. for 22 h. Purification by silica plug (4:2:0.3chloroform:hexane:triethylamine) afforded a yellow-green solid (0.018 g,76% yield). ¹H NMR (400 MHz, CDCl₃) δ 8.77 (bs, 1H, 8.75 (bs, 1H,—N═CH—), 7.82 (bs, 1H, —N═CH—), 7.81 (bs, 1H, —N═CH—), 7.78-7.54 (m, 4H,ArH), 7.25 (bs, 4H, ArH), 5.60 (bs, 4H, CHNHC(O)Ar), 4.08-3.5 (m, 4H,ArOCH₂CH₂—; m, 4H, CH₂C(O)OCH₂(CH₂)₁₄CH₃; m, 4H cyclohexane; m, 4H,ArOCH₂CH₂—), 3.63 (bs, 4H, ArOCH₂CH₂OCH₂CH₂OCH₂CH₂OAr), 3.54 (bs, 4H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 2.36 (m, 4H, cyclohexane), 2.00, 2.35, 2.20, (m,7H, cyclohexane), 1.95, 1.92 (m, s 11H, cyclohexane), 1.61 (m, 4HCH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃), 1.28 (m, 52H, CH₂C(O)OCH₂CH₂(CH₂)₁₃CH₃),1.20-1.00 (m, 10H, cyclohexane), 0.89 (m, 6H, ³J=6.6 HzCH₂C(O)OCH₂(CH₂)₁₄CH₃); MALDITOFMS (MH+)=2222 m/z, (MH+H₂0)=2240 m/z,(MH+2H₂0)=2258 m/z. (MH+3H₂0)=2266 m/z. (MH⁺ 3 dihydroxybenzoic acidmatrix)=2680 m/z.

Example 3

Bridged macrocyclic module compound 13 is prepared according to astepwise synthesis method as shown in Scheme 17.

Example 4

Bridged macrocyclic module 14 is prepared by a convergent synthesismethod as shown in Schemes 18 and 19.

The N-protected synthon trimer A is prepared as shown above in Scheme18:

N-protected synthon trimer A is converted to the activated synthontrimer A, which is combined with bridged program director compound B toproduce bridged macrocyclic module compound 14.

Example 5

Bridged Macrocyclic Module 15 is prepared according to Scheme 20.

Example 6

Transport properties for bridged macrocyclic module 12 (Octamer IV pjs).The dimensions of macrocyclic module and bridged macrocyclic modulepores may be measured by electrical conductance in a voltage-clampedlipid bilayer test. Bridged macrocyclic modules 12 were dissolved into aphosphatidylcholine-phosphatidylethanolamine lipid bilayer. On one sideof the bilayer was placed a solution containing a test cationic species.On the other side was placed a solution containing a cationic speciesknown to be able to pass through the module pore. Anions required forcharge neutrality were selected such that they would not pass throughthe module pore (Cl⁻ in this example). When a positive potential wascreated in the solution on the side of the lipid bilayer containing thetest species, if the test cations were of such a size that they couldnot pass through the pores in the modules, no current would be detected.The voltage was then reversed to create a positive potential on the sideof the lipid bilayer having the solution containing the cationic speciesknown to be able to traverse the pore. Observation of the expectedcurrent confirmed the integrity of the lipid bilayer and theavailability of the module pores as transporters of cations of the knownsize and smaller.

The selective permeability of bridged macrocyclic module 12 (Octamer IVpjs) is shown in Table 9. “Yes” indicates permeation of the solute, “No”indicates rejection of the solute. Permeation and rejection areindicators of clearance. The clamp voltage was 50 mV. Transportproperties of two hexameric macrocyclic modules are shown forcomparison.

TABLE 9 Transport properties for bridged and non-bridged macrocyclicmodules Bridged Solute macro- Hydra- Hexamer I_(a) Hexamer I_(pa) cycliction (1,2-imine) (1,2-amine) module 12 Solute Radius Radius Radius(amide- Solute Radius (2^(nd) shell) 3.3 Å 3.9 Å imine) Li⁺ 0.6 2.0(5.6) No Yes Yes Na⁺ 1.0 2.2 Yes Yes Yes K⁺ 1.3 2.7 Yes Yes Yes Ca²⁺ 1.02.7 Yes Yes No- Blocker Mg²⁺ 0.7 2.8 (5.5) No Yes No NH₃ ⁺ 1.9 2.9 YesYes Yes Cs⁺ 1.7 3 Yes Yes Yes MeNH₃ ⁺ 2 3 Yes Yes No EtNH₃ ⁺ 2.6 3.6 NoYes No NMe₄ ⁺ 2.6 3.6 No Yes No Åmino- 3.1 4.1 No Yes No guanidineCholine 3.8 4.8 No Yes No NEt₄ ⁺ 3.9 4.4 No No No Gluco- 4.2 5.2 No NoNo samine NPr₄ ⁺ — — — No —

In Table 9, “No-blocker” indicates that the ion did not traverse themodule, but was trapped in the module. The results in Table 9 show thatthe cut-off for passage through the pores in the selected bridgedmacrocyclic module is a van der Waals radius of between about 1.8-2.Without wishing to be bound by theory, and recognizing that severalfactors influence pore transport, the observed ability of hydrated ionsto pass through the pore may be due to partial dehydration of thespecies to enter the pore, transport of water molecules and ions throughthe pore separately or with reduced interaction during transport, andre-coordination of water molecules and ions after transport. The detailsof pore structure, composition, and chemistry, the flexibility of themodule, and other interactions may affect the transport process. Withoutwishing to be bound by theory, Mg²⁺ and Ca²⁺ ions may be bound by thePEG bridge moiety in bridged macrocyclic module 12.

Example 7

Preparation of nanofilm of Octamer IV pjs. 25 μl of a 1 mg/ml solutionof Octamer IV pjs in chloroform was spread on a 100 mM NaH₂PO₄ bufferedsubphase (pH 7.0, T=22° C.). After waiting for 19 minutes to allow forspreading solvent evaporation, the monolayer was compressed at 1 mN/m.The monolayer exhibited a collapse pressure of approximately 30 mN/m andthe entire isotherm indicated that the module was highly compressible.The corresponding composite isotherm data (i.e., from several Langmuirfilm runs) is shown in FIG. 1.

The inflection point at approximately 50 Å indicates that the moleculeoccupies a much smaller space per molecule than expected. This was morethan half of the expected area from molecular mechanics computations.

Example 8

Brewster Angle Microscopy (BAM) evaluation of nanofilms from Octamer IVpjs. Nanofilms produced on a Langmuir trough interface at differentsurface pressures were evaluated by BAM, as shown in FIG. 2. In FIG. 2,SP indicates surface pressure. Thus, at moderate surface pressures, theOctamer IV pjs appears to make a homogenous film on the Langmuir trough.

Example 9

Quantum mechanical (QM) and molecular mechanical (MM) computations forapproximation of module pore size. Without intending to be bound by anyone particular theory, one method to approximate pore size of amacrocyclic module or bridged macrocyclic module is quantum mechanical(QM) and molecular mechanical (MM) computations. For the purposes of QMand MM computations, the root mean square deviations in the pore areaswere computed over dynamic runs.

For QM, each module was first optimized using the MM+ force fieldapproach of Allinger (JACS, 1977, 99:8127) and Burkert, et al.,(Molecular Mechanics, ACS Monograph 177, 1982). They were thenre-optimized using the AM1 Hamiltonian (Dewar, et al., JACS, 1985,107:3903; Dewar, et al., JACS, 1986, 108:8075; Stewart, J. Comp. AidedMol. Design, 1990, 4:1). To verify the nature of the potential energysurface in the vicinity of the optimized structures, the associatedHessian matrices were computed using numerical double-differencing.

For MM, the OPLS-AA force field approach (Jorgensen, et al., JACS, 1996,118:11225) was used. For imine linkages, the dihedral angle was confinedto 180°±10°. The structures were minimized and equilibrated for onepicosecond using 0.5 femtosecond time steps. Then a 5 nanoseconddynamics run was carried out with a 1.5 femtosecond time step.Structures were saved every picosecond.

While not wishing to be bound by theory, the molecular dynamics data forOctamer IV pjs indicated that there is a bending effect along the axisof the bridge moiety (FIG. 3), which may account for the unexpectedlylow molecular area observed in the Langmuir isotherm.

Molecular mechanics computations further indicated that the moleculewill bend along the axis of the bridge moiety in solvents of lowdielectric constant. The temperature was set at 300 K for thissimulation. FIGS. 4A and 4B show the molecule before and after thesimulation, with the indicated distances between atoms monitored.

A further molecular mechanics simulation included an annealing andcooling routine in a vacuum. Octamer IV pjs and a identical modulewithout the bridge moiety were subject to 1 ps simulated annealing(T=1000K to 298K) in 10K temperature steps, and subsequently, 5 psrunning at T=298K, and finally, 1 ps simulated annealing (T-298K to 0K)in 10K temperature steps. FIGS. 5A and 5B show the structures before andafter the annealing routine (with optimization performed after themolecular dynamics).

Example 10

Derivatization of SiO₂ substrates withmethylacryloxymethyltrimethoxysilane (MAOMTMOS). SiO₂ substrates werefirst sonicated in a piranha solution (3:1 ratio of H₂SO₄:30% H₂O₂) for15 minutes, followed by a 15 minute sonication in Milli-Q water (>18MΩ-cm). The derivatization step was done in a glove bag under N₂atmosphere. 0.05 mL MAOMTMOS and 0.05 mL pyridine were added to 9 mL oftoluene. Immediately following mixing, the freshly cleaned SiO₂substrates were immersed in the MAOMTMOS solution for 10 min. Substrateswere washed with copious amounts of toluene and then dried with N₂.Deposited MAOMTMOS films showed a range of thickness values from 0.8 to1.3 nm.

Example 11

Deposition of Octamer IV pjs-acrylamide nanofilm on MAOMTMOS modifiedSiO₂ substrates.

Octamer IV pjs-acrylamide is synthesized by addition of acrylamide toOctamer IV pjs.

The derivatized SiO₂ substrates are lowered into a pH 5, 22° C. aqueoussubphase. 170 μl of Octamer IV pjs-acrylamide (1 mg/mL CHCl₃ solution)is spread at the air/water interface of a Langmuir trough. After 10 minthe film is compressed to 5 mN/m at a rate of 2 mm/min. Prior to filmcollapse, the substrates are raised out of the subphase at a rate of 2mm/min, resulting in the deposition of one layer of Octamer IVpjs-acrylamide. Following deposition some samples are irradiated (254nm) for 40 or 220 min to induce coupling between the surface acrylgroups (MAOMTMOS) and the acrylamide group of Octamer IV pjs-acrylamide.Samples are sonicated in CHCl₃ following the UV cure to determine theextent of surface attachment. If the film did not react with thesurface, this treatment should result in the removal of the film.

Example 12

Selective filtration and relative clearance of solutes by variousnanofilms which are contemplated to be produced by compounds of theinvention. In Table 10, the heading “high permeability” indicates aclearance of greater than about 70-90% of the solute. The heading“medium permeability” indicates a clearance of less than about 50-70% ofthe solute. The heading “low permeability” indicates a clearance of lessthan about 10-30% of the solute.

TABLE 10 Clearance of solutes by nanofilms high medium low Nanofilmpermeability permeability permeability water H₂O Glucose, Na⁺, K⁺, Ca²⁺,Mg²⁺, Li⁺, nanofilm phosphate urea, creatinine ion H₂O, Na⁺, K⁺, GlucoseCa²⁺, Mg²⁺, Li⁺, nanofilm phosphate urea, creatinine glucose H₂O, Na⁺,K⁺, Phosphate Ca²⁺, Mg²⁺, Li⁺, nanofilm Glucose urea, creatinine G H₂O,Na⁺, K⁺, Vitamin B₁₂, Myglobin, nanofilm phosphate, Glucose, Insulin, β₂Ovalbumin, Ca²⁺, Mg²⁺, Li⁺, Microglobulin Albumin, urea, creatinine gasHe, H₂ — H₂O and larger, nanofilm liquids in general anion Cl⁻ HCO₃ ⁻,Phosphate — nanofilm

Example 13

Nanofilm filtration function of various nanofilms. The approximatediameter of some various species to be considered in a filtrationprocess are illustrated in Table 11.

TABLE 11 Size of various species of interest for filtration solutemolecular weight (Da) diameter (Å) virus 10⁶ 133 immunoglobulin G (IgG)10⁵ 60 albumin 50 × 10⁴ 50 β₂-Microglobulin 10³ 13 urea 60  — Na⁺ 23  —

The filtration function of a membrane may be described in terms of itssolute rejection profile. The filtration function of some nanofilmmembranes which are contemplated to be produced by the compounds of theinvention are exemplified in Tables 12-13.

TABLE 12 Example filtration function of a G-membrane MOLECULAR SOLUTEWEIGHT PASS/NO PASS Albumin 68 kDa NP Ovalbumin 44 kDa P Myoglobin 17kDa P β₂-Microglobulin 12 kDa P Insulin 5.2 kDa P Vitamin B₁₂ 1350 Da PUrea, H₂O, ions <1000 Da P

TABLE 13 Example filtration function of a T-membrane MOLECULAR SOLUTEWEIGHT PASS/NO PASS β₂-Microglobulin 12 kDa NP Insulin 5.2 kDa NPVitamin B₁₂ 1350 Da NP Glucose 180 Da NP Creatinine 131 Da NP H₂PO₄ ⁻,HPO₄ ²⁻ ≈97 Da NP HCO₃ ⁻ 61 Da NP Urea 60 Da NP K+ 39 Da P Na+ 23 Da P

The passage or exclusion of a solute is measured by its clearance, whichreflects the portion of solute that actually passes through themembrane. The no pass symbol in Tables 12-13 indicates that the soluteis partly excluded by the nanofilm, sometimes less than 90% rejection,often at least 90% rejection, sometimes at least 98% rejection. The passsymbol indicates that the solute is partly cleared by the nanofilm,sometimes less than 90% clearance, often at least 90% clearance,sometimes at least 98% clearance.

Synthon and Macrocyclic Module Synthesis Methods

All chemical structures illustrated and described in this specification,both in the description above and the examples below, as well as in thefigures, are intended to encompass and include all variations andisomers of the structure which are foreseeable, including allstereoisomers and constitutional or configurational isomers when theillustration, description, or figure is not explicitly limited to anyparticular isomer.

Methods for Preparing Cyclic Synthons

To avoid the need to separate single configurational or enantiomericisomers from complex mixtures resulting from non-specific reactions,stereospecific or at least stereoselective coupling reactions may beemployed in the preparation of the synthons of this invention. Thefollowing are examples of synthetic schemes for several classes ofsynthons useful in the preparation of macrocyclic modules of thisinvention. In general, the core synthons are illustrated, and lipophilicmoieties are not shown on the structures, however, it is understood thatall of the following synthetic schemes might encompass additionallipophilic or hydrophilic moieties used to prepare amphiphilic and othermodified macrocyclic modules. Species are numbered in relation to thescheme in which they appear; for example, “S21-1” refers to thestructure 1 in Scheme 21.

An approach to preparing synthons of 1,3-Diaminocyclohex-5-ene is shownin Scheme 21. Enzymatically assisted partial hydrolysis of the

symmetrical diester S21-1 is used to give enantiomerically pure S21-2.S21-2 is subjected to the Curtius reaction and then quenched with benzylalcohol to give protected amino acid S21-3. Iodolactonization ofcarboxylic acid S21-4 followed by dehydrohalogenation gives unsaturatedlactone S21-6. Opening of the lactone ring with sodium methoxide givesalcohol S21-7, which is converted with inversion of configuration toS21-8 in a one-pot reaction involving mesylation, SN₂ displacement withazide, reduction and protection of the resulting amine withdi-tert-butyl dicarbonate. Epimerization of S21-8 to the more stablediequatorial configuration followed by saponification gives carboxylicacid S21-10. S21-10 is subjected to the Curtius reaction. A mixedanhydride is prepared using ethyl chloroformate followed by reactionwith aqueous NaN₃ to give the acyl azide, which is thermally rearrangedto the isocyanate in refluxing benzene. The isocyanate is quenched with2-trimethylsilylethanol to give differentially protected tricarbamateS21-11. Reaction with trifluoroacetic acid (TFA) selectively deprotectsthe 1,3-diamino groups to provide the desired synthon S21-12.

In another variation, an approach to preparing synthons of1,3-Diaminocyclohexane is shown in Scheme 21a.

Some aspects of these preparations are given in Suami et al., J. Org.Chem. 1975, 40, 456 and Kavadias et al. Can. J. Chem. 1978, 56, 404.

In another variation, an approach to preparing synthons of1,3-substituted cyclohexane is shown in Scheme 21b.

This synthon will remain “Z-protected” until the macrocyclic module hasbeen cyclized. Subsequent deprotection to yield a macrocyclic modulewith amine functional groups is done by a hydrogenation protocol.

Norbornanes (bicycloheptanes) may be used to prepare synthons of thisinvention, and stereochemically controlled multifunctionalization ofnorbornanes can be achieved. For example, Diels-Alder cycloaddition maybe used to form norbornanes incorporating various functional groupshaving specific, predictable stereochemistry. Enantiomerically enhancedproducts may also be obtained through the use of appropriate reagents,thus limiting the need for chiral separations.

An approach to preparing synthons of 1,2-Diaminonorbornane is shown inScheme 22. 5-(Benzyloxy-methyl)-1,3-cyclopentadiene (S22-13) is reactedwith

diethylaluminum chloride Lewis acid complex of di-(l-menthyl fumarate(S22-14) at low temperature to give the diastereomerically purenorbornene S22-15. Saponification with potassium hydroxide in aqueousethanol gives the diacid S22-16, which is subjected to a tandem Curtiusreaction with diphenylphosphoryl azide (DPPA), the reaction product isquenched with 2-trimethylsilylethanol to give the biscarbamate S22-17.Deprotection with TFA gives diamine S22-18.

Another approach to this synthon class is outlined in Scheme 23. Openingof anhydride S23-19 with methanol in the presence of quinidine gives theenantiomerically pure ester acid S23-20. Epimerization of the estergroup with sodium methoxide (NaOMe) gives S23-21. A Curtius reactionwith DPPA followed by quenching with trimethylsilylethanol givescarbamate S23-22. Saponification with NaOH gives the acid S23-23, whichundergoes a Curtius reaction,

then quenched with benzyl alcohol to give differentially protectedbiscarbamate S23-24. Compound S23-24 can be fully deprotected to providethe diamine or either of the carbamates can be selectively deprotected.

An approach to preparing synthons of endo,endo-1,3-Diaminonorbornane isshown in Scheme 24. 5-Trimethylsilyl-1,3-cyclopentadiene (S24-25) isreacted with the diethylaluminum chloride Lewis acid complex ofdi-(l)-menthyl fumarate at low temperature to give nearlydiastereomerically pure norbornene S24-26. Crystallization of S24-26from alcohol results in recovery of greater than 99% of the singlediastereomer. Bromolactonization followed by silver mediatedrearrangement gives mixed diester S24-28 with an alcohol moiety at the7-position. Protection of the alcohol with benzyl bromide and selectivedeprotection of the methyl ester gives the free carboxylic acid S24-30.A Curtius reaction results in trimethylsilylethyl carbamate norborneneS24-31. Biscarbonylation of the olefin in methanol, followed by asingle-step deprotection and dehydration gives the mono-anhydrideS24-33. Quinidine mediated opening of the anhydride with methanol givesS24-34. Curtius transformation of S24-34 gives the biscarbamate S24-35,which is deprotected with TFA or tetrabutylammonium fluoride (TBAF) togive diamine S24-36.

Another approach to this class of synthons is outlined in Scheme 25.Benzyl alcohol opening of S23-19 in the presence of quinidine givesS25-37 in high enantiomeric excess. Iodolactonization followed by NaBH₄reduction gives lactone S25-39. Treatment with NaOMe liberates themethyl ester and the free alcohol to generate S25-40. Transformation ofthe alcohol S25-40 to the inverted t-butyl carbamate protected amineS25-41 is accomplished in a one-pot reaction by azide deplacement of themesylate S25-40 followed by reduction to the amine, which is protectedwith di-tert-butyl dicarbonate. Hydrogenolytic cleavage of the benzylester and epimerization of the methyl ester to the exo configuration isfollowed by protection of the free acid with benzyl bromide to giveS25-44. Saponification of the methyl ester followed by atrimethylsilylethanol quenched Curtius reaction

gives the biscarbamate S25-46, which is cleaved with TFA to give thedesired diamine S25-47.

An approach to preparing synthons of exo,endo-1,3-Diaminonorbornane isshown in Scheme 26. p-Methoxybenzyl alcohol opening of norborneneanhydride S23-19 in the presence of quinidine gives monoester S26-48 inhigh enantiomeric excess. Curtius reaction of the free acid givesprotected all endo monoacid-monoamine S26-49. Biscarbonylation andanhydride formation gives exo-monoanhydride S26-51. Selectivemethanolysis in the presence of quinine gives S26-52. Atrimethylsilylethanol quenched Curtius reaction gives biscarbamateS26-53. Epimerization of the two esters results in the more stericallystable S26-54. Cleavage of the carbamate groups provides synthon S26-55.

Methods for Preparing Macrocyclic Modules

Synthons may be coupled to one another to form macrocyclic modules. Inone variation, the coupling of synthons may be accomplished in aconcerted scheme. Preparation of a macrocyclic module by the concertedroute may be performed using, for example, at least two types ofsynthons, each type having at least two functional groups for couplingto other synthons. The functional groups may be selected so that afunctional group of one type of synthon can couple only to a functionalgroup of the other type of synthon. When two types of synthons are used,a macrocyclic module may be formed having alternating synthons ofdifferent types. Scheme 27 illustrates a concerted module synthesis.

Referring to Scheme 27, 1,2-Diaminocyclohexane, S27-1, is a synthonhaving two amino functional groups for coupling to other synthons, and2,6-diformyl-4-dodec-1-ynylphenol, S27-2, is a synthon having two formylgroups for coupling to other synthons. An amino group may couple with aformyl group to form an imine linkage. In Scheme 27, a concerted producthexamer macrocyclic module is shown.

In one variation, a mixture of tetramer, hexamer, and octamermacrocyclic modules may be formed in the concerted scheme. The yields ofthese macrocyclic modules can be varied by changing the concentration ofvarious synthons in the reagent mixture, and among other factors, bychanging the solvent, temperature, and reaction time.

The imine groups of S27-3 can be reduced, e.g. with sodium borohydride,to give amine linkages. If the reaction is carried out using2,6-di(chlorocarbonyl)-4-dodec-1-ynylphenol instead of2,6-diformyl-4-dodec-1-ynylphenol, the resulting module will containamide linkages. Similarly, if 1,2-dihydroxycyclohexane is reacted with2,6-di(chlorocarbonyl)-4-dodec-1-ynylphenol, the resulting module willcontain ester linkages.

In some variations, the coupling of synthons may be accomplished in astepwise scheme. In an example of the stepwise preparation ofmacrocyclic modules, a first type of synthon is substituted with oneprotected functional group and one unprotected functional group. Asecond type of synthon is substituted with an unprotected functionalgroup that will couple with the unprotected functional group on thefirst synthon. The product of contacting the first type of synthon withthe second type of synthon may be a dimer, which is made of two coupledsynthons. The second synthon may also be substituted with anotherfunctional group which is either protected, or which does not couplewith the first synthon when the dimer is formed. The dimer may beisolated and purified, or the preparation may proceed as a one-potmethod. The dimer may be contacted with a third synthon having twofunctional groups, only one of which may couple with the remainingfunctional group of either the first or second synthons to form atrimer, which is made of three coupled synthons. Such stepwise couplingof synthons may be repeated to form macrocyclic modules of various ringsizes. To cyclize or close the ring of the macrocyclic module, then^(th) synthon which was coupled to the product may be substituted witha second functional group which may couple with the second functionalgroup of a previously coupled synthon that has not been coupled, whichmay be deprotected for that step. The stepwise method may be carried outwith synthons on solid phase support. Scheme 28 illustrates a stepwisepreparation of module S28-1.

Compound S28-2 is reacted with S28-3, in which the phenol is protectedas the benzyl ether and the nitrogen is shown as protected with a group“P,” which can be any of a large number of protecting groups well-knownin the art, in the presence of methanesulfonyl chloride (Endo, K.;Takahashi, H. Heterocycles, 1999, 51, 337), to give S28-4. Removal ofthe N-protecting group give the free amine S28-5, which can be coupledwith synthon S28-6 using any standard peptide coupling reaction such asBOP/HOBt to give S28-7. Deprotection/coupling is repeated, alternatingsynthons S28-3. and S28-6 until a linear construct with eight residuesis obtained. The remaining acid and amine protecting groups on the 8-merare removed and the oligomer is cyclized, see e.g., Caba, J. M., et al.,J. Org. Chem., 2001, 66:7568 (PyAOP cyclization) and Tarver, J. E. etal., J. Org. Chem., 2001, 66:7575 (active ester cyclization). The Rgroup is H or an alkyl group coupled via a functional group to thebenzene ring, and X is N, O, or S. Examples of solid supports includeWang resin, hydrogels, silica gels, sepharose, sephadex, agarose, andinorganic solids. Using a solid support might simplify the procedure byobviating purification of intermediates along the way. The finalcyclization may be done in a solid phase mode. A “safety-catch linker”approach (Bourne, G. T., et al., J. Org. Chem., 2001, 66:7706) may beused to obtain cyclization and resin cleavage in a single operation.

In another variation, a concerted method involves contacting two or moredifferent synthons and a linker molecule as shown in Scheme 29, where Rmay be an alkyl group or other lipophilic group.

In another variation, a stepwise linear method involves various synthonsand a solid phase support as shown in Scheme 30.

In another variation, a stepwise convergent method involves synthontrimers and a solid phase support as shown in Scheme 31. This method canalso be done without the solid phase support using trimers in solution.

In another variation, a template method involves synthons broughttogether by a template as shown in Scheme 32. Some aspects of thisapproach (and an Mg²⁺ template) are given in Dutta et al. Inorg. Chem.1998, 37, 5029.

In another variation, a linker molecule method involves cyclizingsynthons in solution as shown in Scheme 33.

Reagents for the following examples were obtained from Aldrich ChemicalCompany and VWR Scientific Products. All reactions were carried outunder nitrogen or argon atmosphere unless otherwise noted. Solventextracts of aqueous solutions were dried over anhydrous Na₂SO₄.Solutions were concentrated under reduced pressure using a rotaryevaporator. Thin layer chromatography (TLC) was done on Analtech Silicagel GF (0.25 mm) plates or on Machery-Nagel Alugram Sil G/UV (0.20 mm)plates. Chromatograms were visualized with either UV light,phosphomolybdic acid, or KMnO₄. All compounds reported were homogenousby TLC unless otherwise noted. HPLC analyses were performed on a HewlettPackard 1100 system using a reverse phase C-18 silica column.Enantiomeric excess was determined by HPLC using a reverse phase(l)-leucine silica column from Regis Technologies. All ¹[H] and ¹³[C]NMR spectra were collected at 400 MHz on a Varian Mercury system.Electrospray mass spectra were obtained by Synpep Corp., or on a ThermoFinnigan LC-MS system.

Example 14 2,6-Diformyl-4-bromophenol

Hexamethylenetetramine (73.84 g, 526 mmol) was added to TFA (240 mL)with stirring. 4-Bromophenol (22.74 g, 131 mmol) was added in oneportion and the solution heated in an oil bath to 120° C. and stirredunder argon for 48 h. The reaction mixture was then cooled to ambienttemperature. Water (160 mL) and 50% aqueous H₂SO₄ (80 mL) were added andthe solution stirred for an additional 2 h. The reaction mixture waspoured into water (1600 mL) and the resulting precipitate collected on aBüchner funnel. The precipitate was dissolved in ethyl acetate (EtOAc)and the solution was dried over MgSO₄. The solution was filtered and thesolvent removed on a rotary evaporator. Purification by columnchromatography on silica gel (400 g) using a gradient of 15-40% ethylacetate in hexanes resulted in a isolation of the product as a yellowsolid (18.0 g, 60%).

¹H NMR (400 MHz, CDCl₃) δ 11.54 (s, 1H, OH), 10.19 (s, 2H, CHO), 8.08(s, 2H, ArH).

Example 15 2,6-Diformyl-4-(dodecyn-1-yl)phenol

2,6-Diformyl-4-bromophenol (2.50 g, 10.9 mmol), 1-dodecyne (2.00 g, 12.0mmol), CuI (65 mg, 0.33 mmol), and bis(triphenylphosphine)palladium)II)dichloride were suspended in degassed acetonitrile (MeCN) (5 mL) anddegassed benzene (1 mL). The yellow suspension was sparged with argonfor 30 min and degassed Et₃N (1 mL) was added. The resulting brownsuspension was sealed in a pressure vial, warmed to 80° C. and heldthere for 12 h. The mixture was then partitioned between EtOAc and KHSO₄solution. The organic layer was separated, washed with brine, dried(MgSO₄) and concentrated under reduced pressure. The dark yellow oil waspurified by column chromatography on silica gel (25% Et₂O in hexanes) togive 1.56 g (46%) of the title compound.

¹H NMR (400 MHz, CDCl₃)

11.64 (s, 1H, OH), 10.19 (s, 2H, CHO), 7.97 (s, 2H, ArH), 2.39 (t, 2H,J=7.2 Hz, propargylic), 1.59 (m, 3H, aliphatic), 1.43, (m, 2H,aliphatic), 1.28 (m, 11H, aliphatic), 0.88 (t, 3H, J=7.0 Hz, CH₃).

¹³C NMR (400 MHz, CDCl₃) δ 192.5, 162.4, 140.3, 122.8, 116.7, 91.4,77.5, 31.9, 29.6, 29.5, 29.3, 29.1, 28.9, 28.5, 22.7, 19.2, 14.1.

MS (FAB): Calcd. for C₂₀H₂₇O₃ 315.1960; found 315.1958 [M+H]⁺.

Example 16 2,6-Diformyl-4-(dodecen-1-yl)phenol

2,6-Diformyl-4-bromophenol (1.00 g, 4.37 mmol), 1-dodecene (4.8 mL, 21.7mmol), 1.40 g tetrabutylammonium bromide (4.34 mmol), 0.50 g NaHCO₃(5.95 mmol), 1.00 g LiCl (23.6 mmol) and 0.100 g palladium diacetate(Pd(OAc)₂) (0.45 mmol) were combined in 30 mL degassed anhydrousdimethylformamide (DMF). The mixture was sparged with argon for 10 minand then sealed in a pressure vial which was warmed to 82° C. and heldfor 40 h. The crude reaction mixture was partitioned between CH₂Cl₂ and0.1 M HCl solution. The organic layer was washed with 0.1 M HCl (2×),brine (2×), and saturated aqueous NaHCO₃ (2×), dried over MgSO₄ andconcentrated under reduced pressure. The dark yellow oil was purified bycolumn chromatography on silica gel (25% hexanes in Et₂O) to give 0.700g (51%) of the title compound as primarily the Z isomer.

¹H NMR (400 MHz, CDCl₃)

11.50 (s, 1H, OH), 10.21 (s, 2H, CHO), 7.95 (s, 2H, ArH), 6.38 (d, 1H,vinyl), 6.25 (m, 1H, vinyl), 2.21 (m, 2H, allylic), 1.30-1.61 (m, 16H,aliphatic), 0.95 (t, 3H, J=7.0 Hz, CH₃).

MS (FAB): Calcd. for C₂₀H₂₇O₃ 315.20; found 315.35 [M−H]⁻.

Example 17 (1R,6S)-6-Methoxycarbonyl-3-cyclohexene-1-carboxylic Acid(S21-2)

S21-1 (15.0 g, 75.7 mmol) was suspended in pH 7 phosphate buffer (950mL). Pig liver esterase (2909 units) was added, and the mixture stirredat ambient temperature for 72 h with the pH maintained at 7 by additionof 2M NaOH. The reaction mixture was washed with ethyl acetate (200 mL),acidified to pH 2 with 2M HCl, and extracted with ethyl acetate (3×200mL). The extracts were combined, dried, and evaporated to afford 13.8 g(99%) of S21-2.

¹H NMR: (CDCl₃) δ 2.32 (dt, 2H, 2_(ax)- and 5_(ax)-H's), 2.55 (dt, 2H,2_(eq)- and 5_(eq)-H's), 3.00 (m, 2H, 1- and 6-H's), 3.62 (s, 3H,CO₂Me), 5.61 (m, 2H, 3- and 4-H's).

Example 18 Methyl(1S,6R)-6-Benzyloxycarbonylaminocyclohex-3-enecarboxylate (S21-3)

S21-2 (10.0 g, 54.3 mmol) was dissolved in benzene (100 mL) under N₂.Triethylamine (13.2 g, 18.2 mL, 130.3 mmol) was added followed by DPPA(14.9 g, 11.7 mL, 54.3 mmol). The solution was refluxed for 20 h. Benzylalcohol (5.9 g, 5.6 mL, 54.3 mmol) was added and reflux continued for 20h. The solution was diluted with EtOAc (200 mL), washed with saturatedaqueous NaHCO₃ (2×50 mL), water (20 mL), and saturated aqueous NaCl (20mL), dried and evaporated to give 13.7 g (87%) of S21-3.

¹H NMR: (CDCl₃) δ 2.19 (dt, 1H, 5_(ax)-H), 2.37 (tt, 2H, 2_(ax)- and5_(eq)-H's), 2.54 (dt, 1H, 2_(eq)-H), 2.82 (m, 1H, 1-H), 3.65 (s, 3H,CO₂Me), 4.28 (m, 1H, 6-H), 5.08 (dd, 2H, CH₂Ar), 5.42 (d, 1H, NH), 5.62(ddt, 2H, 3- and 4-H's), 7.35 (m, 5H, Ar H's).

Example 19 (1S,6R)-6-Benzyloxycarbonylaminocyclohex-3-enecarboxylic acid(S21-4)

S21-3 (23.5 g, 81.3 mmol) was dissolved in MeOH (150 mL) and thesolution cooled to 0° C. 2M NaOH (204 mL, 0.41 mol) was added, themixture allowed to come to ambient temperature and then it was stirredfor 48 h. The reaction mixture was diluted with water (300 mL),acidified with 2M HCl, and extracted with dichloromethane (250 mL),dried, and evaporated. The residue was recrystallized from diethyl etherto give 21.7 (97%) of S21-4.

¹H NMR: (CDCl₃) δ 2.20 (d, 1H, 5_(ax)-H), 2.37 (d, 2H, 2_(ax)- and5_(eq)-H's), 2.54 (d, 1H, 2_(eq)-H), 2.90 (br s, 1H, 1-H), 4.24 (br s,1H, 6-H), 5.08 (dd, 2H, CH₂Ar), 5.48 (d, 1H, NH), 5.62 (dd, 2H, 3- and4-H's), 7.35 (m, 5H, Ar H's).

Example 20(1S,2R,4R,5R)-2-Benzyloxycarbonylamino-4-iodo-7-oxo-6-oxabicyclo[3.2.1]octane(S21-5)

S21-4 (13.9 g, 50.5 mmol) was dissolved in dichloromethane (100 mL)under N₂, 0.5 M NaHCO₃ (300 mL), KI (50.3 g, 303.3 mmol), and iodine(25.6 g, 101 mmol) were added and the mixture stirred at ambienttemperature for 72 h. The mixture was diluted with dichloromethane (50mL) and the organic phase separated. The organic phase was washed withsaturated aqueous Na₂S₂O₃ (2×50 mL), water (30 mL), and saturatedaqueous NaCl (20 mL), dried and evaporated to afford 16.3 g (80%) ofS21-5.

¹H NMR: (CDCl₃) δ 2.15 (m, 1H, 8_(ax)-H), 2.42 (m, 2H, 3_(ax)- and8_(eq)-H's), 2.75 (m, 2H, 1- and 3_(eq)-H's), 4.12 (br s, 1H, 2-H), 4.41(t, 1H, 4-H), 4.76 (dd, 1H, 5-H), 4.92 (d, 1H, NH), 5.08 (dd, 2H,CH₂Ar), 7.35 (m, 5H, Ar H's).

Example 21(1S,2R,5R)-2-Benzyloxycarbonylamino-7-oxo-6-oxabicyclo[3.2.1]oct-3-ene(S21-6)

S21-5 (4.0 g, 10 mmol) was dissolved in benzene (50 mL) under N₂.1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (1.8 g, 12 mmol) was added andthe solution refluxed for 16 h. The precipitate was filtered and thefiltrate was diluted with EtOAc (200 mL). The filtrate was washed with1M HCl (20 mL), saturated aqueous Na₂S₂O₃ (20 mL), water (20 mL), andsaturated aqueous NaCl (20 mL), dried and evaporated to give 2.2 g (81%)S21-6.

¹H NMR: (CDCl₃) δ 2.18 (d, 1H, 8_(ax)-H), 2.39 (m, 1H, 8_(eq)-H), 3.04(t, 1H, 1-H), 4.70 (m, 1H, 5-H), 4.82 (t, 1H, 2-H), 5.15 (dd, 3H, CH₂Arand NH), 5.76 (d, 1H, 4-H), 5.92 (m, 1H, 3-H), 7.36 (s, 5H, Ar H's).

Example 22 (1S,2R,5R)-Methyl2-Benzyloxycarbonylamino-5-hydroxycyclohex-3-enecarboxylate (S21-7)

S21-6 (9.0 g, 33 mmol) was suspended in MeOH (90 mL) and cooled to 0° C.NaOMe (2.8 g, 52.7 mmol) was added and the mixture stirred for 3 hduring which time a solution gradually formed. The solution wasneutralized with 2M HCl, diluted with saturated aqueous NaCl (200 mL),and extracted with dichloromethane (2×100 mL). The extracts werecombined, washed with water (20 mL) and saturated aqueous NaCl (20 ml),dried, and evaporated. The residue was flash chromatographed (silica gel(250 g), 50:50 hexane/EtOAc) to give 8.5 g (85%) of S21-7.

¹H NMR: (CDCl₃) δ 1.90 (m, 1H, 6_(ax)-H), 2.09 (m, 1H, 6_(eq)-H), 2.81(m, 1H, 1-H), 3.55 (s, 3H, CO₂Me), 4.15 (m, 1H, 5-H), 4.48 (t, 1H, 2-H),5.02 (dd, 2H, CH₂Ar), 5.32 (d, 1H, NH), 5.64 (dt, 1H, 4-H), 5.82 (dt,1H, 3-H), 7.28 (s, 5H, Ar H's).

Example 23 (1S,2R,5S)-Methyl2-Benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylate(S21-8)

S21-7 (7.9 g, 25.9 mmol) was dissolved in dichloromethane (150 mL) andcooled to 0° C. under N₂. Triethylamine (6.3 g, 8.7 mL, 62.1 mmol) andmethanesulfonyl chloride (7.1 g, 62.1 mmol) were added and the mixturestirred at 0° C. for 2 h. (n-Bu)₄NN₃ (14.7 g, 51.7 mmol) indichloromethane (50 mL) was added and stirring continued at 0° C. for 3h followed by 15 h at ambient temperature. The mixture was cooled to 0°C. and P(n-Bu)₃ (15.7 g, 19.3 mL, 77.7 mmol) and water (1 mL) were addedand the mixture stirred at ambient temperature for 24 h. Di-tert-butyldicarbonate (17.0 g, 77.7 mmol) was added and stirring continued for 24h. The solvent was removed, the residue dissolved in 2:1 hexane/EtOAc(100 mL), the solution filtered, and evaporated. The residue was flashchromatographed (silica gel (240 g), 67:33 hexane/EtOAc) to give 5.9 g(56%) of S21-8.

¹H NMR: (CDCl₃) δ 1.40 (s, 9H, Boc H's), 1.88 (m, 1H, 6_(ax)-H), 2.21(m, 1H, 6_(eq)-H), 2.95 (m, 1H, 1-H), 3.60 (s, 3H, CO₂Me), 4.15 (d, 1H,Boc NH), 4.50 (m, 2H, 2- and 5-H's), 5.02 (s, 2H, CH₂Ar), 5.38 (d, 1H, ZNH), 5.65 (m, 2H, 3- and 4-H's), 7.30 (s, 5H, Ar H's).

Example 24 (1R,2R,5S)-Methyl2-Benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylate(S21-9)

S21-8 (1.1 g, 2.7 mmol) was suspended in MeOH (50 mL). NaOMe (0.73 g,13.6 mmol) was added and the mixture refluxed for 18 h after which 0.5 MNH₄Cl (50 mL) was added and the resulting precipitate collected. Thefiltrate was evaporated and the residue triturated with water (25 mL).The insoluble portion was collected and combined with the originalprecipitate to give 0.85 g (77%) of S21-9.

¹H NMR: (CDCl₃) δ 1.38 (s, 9H, Boc H's), 1.66 (m, 1H, 6_(ax)-H), 2.22(d, 1H, 6_(eq)-H), 2.58 (t, 1H, 1-H), 3.59 (3, 3H, CO₂Me), 4.22 (br s,1H, Boc NH), 4.50 (m, 2H, 2- and 5-H's), 4.75 (d, 1H, Z NH), 5.02 (s,2H, CH₂Ar), 5.62 (s, 2H, 3- and 4-H's), 7.30 (s, 5H, Ar H's).

Example 25(1R,2R,5S)-2-Benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylicacid (S21-10)

S21-9 (0.85 g, 2.1 mmol) was suspended in 50:50 MeOH/dichloromethane (5mL) and cooled to 0° C. under N₂ after which 2M NaOH (2.0 mL) was addedand the mixture stirred at ambient temperature for 16 h. The mixture wasacidified with 2M HCl upon which a white precipitate formed. Theprecipitate was collected, washed with water and hexane, and dried togive 0.74 g (90%) of S21-10.

¹H NMR: (CD₃OD) δ 1.42 (s, 9H, Boc H's), 1.66 (m, 1H, 6_(ax)-H), 2.22(d, 1H, 6_(eq)-H), 2.65 (t, 1H, 1-H), 4.18 (m, 1H, 5-H), 4.45 (m, 1H,5-H), 5.04 (s, 2H, CH₂Ar), 5.58 (m, 2H, 3- and 4-H's), 7.35 (s, 5H, ArH's).

Example 26(1R,2R,5S)-2-Benzyloxycarbonylamino-5-t-butoxycarbonylamino-1-(2-trimethylsilyl)ethoxycarbonylaminocyclohex-3-ene(S21-11)

S21-10 (3.1 g, 7.9 mmol) was dissolved in THF (30 mL) under N₂ andcooled to 0° C. Triethylamine (1.6 g, 2.2 mL, 15.9 mmol) was addedfollowed by ethyl chloroformate (1.3 g, 1.5 mL, 11.8 mmol). The mixturewas stirred at 0° C. for 1 h. A solution of NaN₃ (1.3 g, 19.7 mmol) inwater (10 mL) was added and stirring at 0° C. was continued for 2 h. Thereaction mixture was partitioned between EtOAc (50 mL) and water (50mL). The organic phase was separated, dried, and evaporated. The residuewas dissolved in benzene (50 mL) and refluxed for 2 h.2-Trimethylsilylethanol (1.0 g, 1.2 mL, 8.7 mmol) was added and refluxcontinued for 3 h. The reaction mixture was diluted with EtOAc (200 mL),washed with saturated aqueous NaHCO₃ (50 mL), water (20 mL), andsaturated aqueous NaCl (20 mL), dried and evaporated. The residue wasflash chromatographed (silica gel (100 g), 67:33 hexane/EtOAc) to give3.1 g (77%) of S21-11.

¹H NMR: (CDCl₃) δ −0.02 (s, 9H, TMS), 0.90 (t, 3H, CH₂TMS), 1.40 (s, 9H,Boc H's), 2.38 (m, 1H, 6_(eq)-H), 3.62 (m, 1H, 1-H), 4.08 (m, 2H,OCH₂CH₂TMS), 4.18 (m, 1H), 4.38 (m, 1H), 4.62 (m, 1H), 5.07 (dd, 2H,CH₂Ar), 5.18 (m, 1H), 5.26 (m, 1H), 5.58 (d, 1H, olefinic H), 5.64 (d,2H, olefinic H), 7.30 (s, 5, Ar H's).

Example 27 (1R,2R,5S)-2-Benzyloxycarbonylamino-1,5-diaminocyclohex-3-ene(S21-12)

S21-11 (2.5 g, 4.9 mmol) was added to TFA (10 mL) and the solutionstirred at ambient temperature for 16 h after which the solution wasevaporated. The residue was dissolved in water (20 mL), basified to pH14 with KOH and extracted with dichloromethane (3×50 mL). The extractswere combined, washed with water (20 mL), dried and evaporated to give1.1 g (85%) of S21-12.

¹H NMR: (CDCl₃) δ 1.30 (m, 1H, 6_(ax)-H), 2.15 (br d, 1H, 6_(eq)-H),2.68 (m, 1H, 1-H), 3.42 (br s, 1H, 5-H), 3.95 (m, 1H, 2-H), 4.85 (d, 1H,Z NH), 5.08 (t, 2H, CH₂Ar), 5.45 (d, 1H, 4-H), 5.62 (d, 1H, 3-H), 7.32(s, 5H, Ar H's). ESCI MS m/e 262 M+1.

Example 28

Isolation of S21b-2 was accomplished using the following procedure:Using Schlenk technique 5.57 g (10.0 mmol) of methyl ester compound,S21b-1, was dissolved in 250 mL of THF. In another flask LiOH (1.21 g,50.5 mmol) was dissolved in 50 mL water and de-gassed by bubbling N₂through the solution using a needle for 20 minutes. The reaction wasstarted transferring the base solution into the flask containing S21b-1over one minute with rapid stirring. The mixture was stirred at roomtemperature and work-up initiated when the starting material S21b-1 wascompletely consumed (Using a solvent system of 66% EtOAc/33% Hexane anddeveloping with phosphomolybdic acid reagent (Aldrich #31, 927-9) thestarting material S21b-1 has an Rf of 0.88 and the product streaks withan Rf of approx. 0.34 to 0.64). The reaction usually takes 2 days.Work-Up: The THF was removed by vacuum transfer until about the samevolume is left as water added to the reaction, in this case 50 mL.During this the reaction solution forms a white mass that adheres to thestir bar surrounded by clear yellow solution. As the THF is beingremoved a separatory funnel is set up including a funnel to pour in thereaction solution and an Erlenmeyer flask is placed underneath theseparatory funnel. Into the Erlenmeyer flask is added some anhydrousNa₂SO₄. This apparatus should be set up before acidification is started.(It is important to set up the separatory funnel and Erlenmeyer flasketc. before acidification of the reaction solution to enable separationof phases and extraction of the product away from the acid quickly oncethe solution attains a pH close to 1. If the separation is not preformedrapidly the Boc functional group will be hydrolyzed significantlyreducing the yield.) Once the volatiles are sufficiently removed, CH₂Cl₂(125 mL) and water (65 mL) are added and the reaction flask cooled in anice bath. The solution is stirred rapidly and 5 mL aliquots of 1N HClare added by syringe and the reaction solution tested with pH paper.Acid is added until the spot on the pH paper shows red (not orange)around the edge indicating a pH is 1 to 2 has been achieved (Thesolution being tested is a mixture of CH₂Cl₂ and water so the pH paperwill show the accurate measurement at the edge of the spot and not thecenter) and the phases are separated by quickly pouring the solutioninto the separatory funnel. As the phases separate the stopcock isturned to release the CH₂Cl₂ phase (bottom) into the Erlenmeyer flaskand swirl the flask to allow the drying agent to absorb water in thesolution. (At this scale of this procedure 80 mL of 1N HCl was used.)Soon after phase separation the aqueous phase is extracted with CH₂Cl₂(2×100 mL), dried over anhydrous Na₂SO₄ and the volatiles removed toproduce 5.37 g/9.91 mmoles of a beautiful white microcrystals reflectinga 99.1% yield. This product can not be purified by chromatography sincethat process would also hydrolyze the Boc functional group on thecolumn.

¹H NMR (400 MHz, CDCl₃)

7.33, 7.25 (5H, m, Ph), 6.30 (1H, d, NH), 5.97 (1H, d, NH), 5.10 (2H, m,CH₂Ph), 4.90 (1H, d, NH), 3.92, 3.58, 3.49 (1H, m, CHNH), 2.96, 2.48,2.04, 1.95, 1.63 (1H, m, CH₂CHNH), 1.34 (9H, s, CCH₃).

IR (crystalline, cm⁻¹) 3326 br w, 3066 w, 3033 w, 2975 w, 2940 w sh,1695 vs, 1506 vs, 1454 m sh, 1391 w, 1367 m, 1300 m sh, 1278 m sh, 1236s, 1213 w sh, 1163 vs, 1100 w, 1053 m, 1020 m, 981 w sh, 910 w, 870 m,846 w, 817 w, 775 w sh, 739 m, 696 m.

Example 29 Di-(l)-menthylbicyclo[2.2.1]hept-5-ene-7-anti-(trimethylsilyl)-2-endo-3-exo-dicarboxylate(S24-26)

To a solution of S24-25 (6.09 g, 0.0155 mol) in toluene (100 mL) wasadded diethylaluminum chloride (8.6 mL of a 1.8 M solution in toluene)at −78° C. under nitrogen and the mixture was stirred for 1 hour. To theresulting orange solution was added S22-14 (7.00 g, 0.0466 mol) dropwiseas a −78° C. solution in toluene (10 mL). The solution was kept at −78°C. for 2 hours, followed by slow warming to room temperature overnight.The aluminum reagent was quenched with a saturated solution of ammoniumchloride (50 mL). The aqueous layer was separated and extracted withmethylene chloride (100 mL) which was subsequently dried over magnesiumsulfate. Evaporation of the solvent left a yellow solid that waspurified by column chromatography (10% ethyl acetate/hexanes) to giveS24-26 as a while solid (7.19 g, 0.0136 mol, 87% yield).

¹H NMR: (CDCl₃) δ −0.09 (s, 9H, SiMe₃), 0.74-1.95 (multiplets, 36H,menthol), 2.72 (d, 1H, α-menthyl carbonyl CH), 3.19 (bs, 1H, bridgeheadCH), 3.30 (bs, 1H, bridgehead CH), 3.40 (t, 1H, α-menthyl carbonyl CH),4.48 (d of t, 1H, α-menthyl ester CH), 4.71 (d of t, 1H, α-menthyl esterCH), 5.92 (d of d, 1H, CH═CH), 6.19 (d of d, 1H, CH═CH).

Example 305-exo-Bromo-3-exo-(l)-menthylcarboxybicyclo[2.2.1]heptane-7-anti-(trimethylsilyl)-2,6-carbolactone(S24-27)

A solution of bromine (3.61 g, 0.0226 mol) in methylene chloride (20 mL)was added to a stirring solution of S24-26 (4.00 g, 0.00754 mol) inmethylene chloride (80 mL). Stirring was continued at room temperatureovernight. The solution was treated with 5% sodium thiosulfate (150 mL),and the organic layer separated and dried over magnesium sulfate. Thesolvent was evaporated at reduced pressure, and the crude productpurified by column chromatography (5% ethyl acetate/hexanes) to giveS24-27 as a white solid (3.53 g, 0.00754 mol, 99% yield).

¹H NMR: (CDCl₃) δ −0.19 (s, 9H, SiMe₃), 0.74-1.91 (multiplets, 18H,menthol), 2.82 (d, 1H, α-lactone carbonyl CH), 3.14 (bs, 1H, lactonebridgehead CH), 3.19 (d of d, 1H, bridgehead CH), 3.29 (t, 1H, α-menthylcarbonyl CH), 3.80 (d, 1H, α-lactone ester), 4.74 (d of t, 1H, α-menthylester CH), 4.94 (d, 1H, bromo CH).

Example 31Bicyclo[2.2.1]hept-5-ene-7-syn-(hydroxy)-2-exo-methyl-3-endo-(l)-menthyldicarboxylate (S24-28)

S24-27 (3.00 g, 0.00638 mol) was dissolved in anhydrous methanol (150mL), silver nitrate (5.40 g, 0.0318 mol) added and the suspensionrefluxed for 3 days. The mixture was cooled, filtered through Celite andthe solvent evaporated to give an oily residue. Purification by columnchromatography gave S24-28 as a light yellow oil (1.72 g, 0.00491 mol,77% yield).

¹H NMR: (CDCl₃) δ 0.75-2.02 (multiplets, 18H, menthol), 2.83 (d, 1H,α-menthyl carbonyl CH), 3.03 (bs, 1H, bridgehead CH), 3.14 (bs, 1H,bridgehead CH), 3.53 (t, 1H, α-methyl carbonyl CH), 3.76 (s, 3H, CH₃),4.62 (d of t, 1H, α-menthyl ester CH), 5.87 (d of d, 1H, CH═CH), 6.23 (dof d, 1H, CH═CH).

Example 322-exo-Methyl-3-endo-(l)-menthylbicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)dicarboxylate (S24-29)

Benzyl bromide (1.20 g, 0.0070 mol) and silver oxide (1.62 g, 0.0070mol) were added to a stirring solution of S24-28 (0.490 g, 0.00140 mol)in DMF (25 mL). The suspension was stirred overnight and then dilutedwith ethyl acetate (100 mL). The solution was washed repeatedly withwater followed by 1 N lithium chloride. The organic layer was separatedand dried with magnesium sulfate. The solvent was evaporated underreduced pressure and the crude product was purified by columnchromatography on silica gel to give S24-29 as an oil (0.220 g, 0.000500mol, 36% yield).

¹H NMR: (CDCl₃) δ 0.74-2.08 (multiplets, 18H, menthol), 2.83 (d, 1H,α-menthyl carbonyl CH), 3.18 (bs, 1H, bridgehead CH), 3.44 (bs, 1H,bridgehead CH), 3.52 (t, 1H, bridge CH), 3.57 (s, 3H, CH₃), 3.68 (t, 1H,α-methyl carbonyl CH), 4.42 (d of d, 2H, benzyl —CH₂—), 4.61 (d of t,1H, α-menthyl ester CH), 5.89 (d of d, 1H, CH═CH), 6.22 (d of d, 1H,CH═CH), 7.25-7.38 (m, 5H, C₆H₅).

Example 33Bicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)-2-exo-carboxy-3-endo-(l)-menthylcarboxylate (S24-30)

S24-29 (0.220 g, 0.00050 mol) was added to a mixture of tetrahydrofuran(1.5 mL), water (0.5 mL), and methanol (0.5 mL). Potassium hydroxide(0.036 g, 0.00065 mol) was added and the solution stirred at roomtemperature overnight. The solvent was evaporated under reduced pressureand the residue purified by column chromatography (10% ethylacetate/hexanes) to give S24-30 (0.050 g, 0.00012 mol, 23% yield).

¹H NMR: (CDCl₃) δ 0.73-2.01 (multiplets, 18H, menthol), 2.85 (d, 1H,α-menthyl carbonyl CH), 3.18 (bs, 1H, bridgehead CH), 3.98 (bs, 1H,bridgehead CH), 3.53 (bs, 1H, bridge CH), 3.66 (t, 1H, α-methyl carbonylCH), 4.44 (d of d, 2H, benzyl —CH₂—), 4.63 (d of t, 1H, α-menthyl esterCH), 5.90 (d of d, 1H, CH═CH), 6.23 (d of d, 1H, CH═CH), 7.25-7.38 (m,5H, C₆H₅).

Mass Spec: calculated for C₂₆H₃₄O₅ 426.24; found 425.4 (M−1) and 851.3(2M−1).

Example 34Bicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthylcarboxylate (S24-31)

To a solution of S24-30 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours thencooled to room temperature. Trimethylsilylethanol is added, and thesolution refluxed for an additional 48 hours. The benzene solution ispartitioned between ethyl acetate and 1 M sodium bicarbonate. Theorganic layers are combined, washed with 1 M sodium bicarbonate anddried over sodium sulfate. The solvent is evaporated under reducedpressure to give the crude Curtius reaction product.

Example 35Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-5-exo-methyl-6-exo-methyltricarboxylate (S24-32)

S24-31, dry copper(II) chloride, 10% Pd/C, and dry methanol are added toa flask with vigorous stirring. After degassing, the flask is chargedwith carbon monoxide to a pressure just above 1 atm., which ismaintained for 72 hours. The solids are filtered and the residue workedup in the usual way to afford the biscarbonylation product.

Example 36Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthylcarbox-5-exo-6-exo-dicarboxylicanhydride (S24-33)

A mixture of S24-32, formic acid, and a catalytic amount ofp-toluenesulfonic acid is stirred at 90° C. overnight. Acetic anhydrideis added and the reaction mixture refluxed for 6 hours. Removal of thesolvents and washing with ether gives the desired anhydride.

Example 37Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-6-exo-carboxy-5-exo-methyldicarboxylate (S24-33)

To a solution of S24-32 in equal amounts of toluene and carbontetrachloride is added quinidine. The suspension is cooled to −65° C.and stirred for 1 hour. Three equivalents of methanol are slowly addedover 30 minutes. The suspension is stirred at −65° C. for 4 daysfollowed by removal of the solvents under reduced pressure. Theresulting white solid is partitioned between ethyl acetate and 2M HCl.The quinine is recovered from the acid layer and S24-33 obtained fromthe organic layer.

Example 38Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-6-exo-(trimethylsilylethoxycarbonyl)amino-5-exo-methyldicarboxylate (S24-35)

To a solution of S24-34 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours. Aftercooling to room temperature, 2-trimethylsilylethanol is added and thesolution refluxed for 48 hours. The benzene solution is partitionedbetween ethyl acetate and 1M sodium bicarbonate. The organic layers arecombined, washed with 1M sodium bicarbonate, and dried over sodiumsulfate. The solvent is evaporated under reduced pressure to give thecrude Curtius reaction product.

Example 39endo-Bicyclo[2.2.1]hept-5-ene-2-benzylcarboxylate-3-carboxylic acid(S25-37)

Compound S23-19 (4.00 g, 0.0244 mol) and quinidine (8.63 g, 0.0266 mol)were suspended in equal amounts of toluene (50 mL) and carbontetrachloride (50 mL). The suspension was cooled to −55° C. after whichbenzyl alcohol (7.90 g, 0.0732 mol) was added over 15 minutes. Thereaction mixture became homogenous after 3 hours and was stirred at −55°C. for an additional 96 hours. After removal of the solvents, theresidue was partitioned between ethyl acetate (300 mL) and 2Mhydrochloric acid (100 mL). The organic layer was washed with water(2×50 mL) and saturated aqueous sodium chloride (1×50 mL). Drying overmagnesium sulfate and evaporation of the solvent gave S25-37 (4.17 g,0.0153 mol, 63% yield).

¹H NMR: (CDCl₃) δ 1.33 (d, 1H, bridge CH₂), 1.48 (d of t, 1H, bridgeCH₂), 3.18 (bs, 1H, bridgehead CH), 3.21 (bs, 1H, bridgehead CH), 3.33(t, 2H, α-acid CH), 4.98 (d of d, 2H, CH₂Ph), 6.22 (d of d, 1H, CH═CH),6.29 (d of d, 1H, CH═CH), 7.30 (m, 5H, C₆H₅).

Example 402-endo-Benzylcarboxy-6-exo-iodobicyclo[2.2.1]heptane-3,5-carbolactone(S25-38)

S25-37 (4.10 g, 0.0151 mol) was dissolved in 0.5 M sodium bicarbonatesolution (120 mL) and cooled to 0° C. Potassium iodide (15.0 g, 0.090mol) and iodine (7.66 g, 0.030 mol) were added followed by methylenechloride (40 mL). The solution was stirred at room temperatureovernight. After dilution with methylene chloride (100 mL), sodiumthiosulfate was added to quench the excess iodine. The organic layer wasseparated and washed with water (100 mL) and sodium chloride solution(100 mL). Drying over magnesium sulfate and evaporation of the solventgave S25-38 (5.44 g, 0.0137 mol, 91% yield).

¹H NMR: (CDCl₃) δ 1.86 (d of q, 1H, bridge —CH₂—), 2.47 (d of t, 1H,bridge —CH₂—), 2.83 (d of d, 1H, α-lactone carbonyl CH), 2.93 (bs, 1H,lactone bridgehead CH), 3.12 (d of d, 1H, α-benzyl ester CH), 3.29 (m,1H, bridgehead CH), 4.63 (d, 1H, α-lactone ester CH), 5.14 (d of d, 2H,CH₂Ph), 5.19 (d, 1H, iodo CH), 7.38 (m, 5H, C₆H₅).

Example 41 2-endo-Benzylcarboxy-bicyclo[2.2.1]heptane-3,5-carbolactone(S25-39)

S25-38 (0.30 g, 0.75 mmol) was placed in DMSO under N₂, NaBH₄ (85 mg,2.25 mmol) added and the solution stirred at 85° C. for 2 h. The mixturewas cooled, diluted with water (50 mL) and extracted withdichloromethane (3×20 mL). The extracts were combined, washed with water(4×15 mL) and saturated aqueous NaCl (10 mL), dried, and evaporated togive 0.14 g (68%) of S25-39.

Example 425-endo-hydroxybicyclo[2.2.1]heptane-2-endo-benzyl-3-endo-methyldicarboxylate (S25-40)

Compound S25-39 is dissolved in methanol and sodium methoxide added withstirring. Removal of the solvent gives S25-40.

Example 43Bicyclo[2.2.1]heptane-2-endo-benzyl-3-endo-methyl-5-exo-(t-butoxycarbonyl)-aminodicarboxylate (S25-41)

In a one-pot reaction S25-40 is converted to the corresponding mesylatewith methanesulfonyl chloride, sodium azide added to displace themesylate to give exo-azide, which is followed by reduction with tributylphosphine to give the free amine, which is protected as the t-Bocderivative to give S25-41.

Example 44Bicyclo[2.2.1]heptane-2-endo-carboxy-3-exo-methyl-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S25-42)

The benzyl ether protecting group is removed by catalytic hydrogenolysisof S25-41 with 10% Pd/C in methanol at room temperature for 6 hours.Filtration of the catalyst and removal of the solvent yields crudeS25-42.

Example 45Bicyclo[2.2.1]heptane-2-endo-carboxy-3-exo-methyl-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S25-43)

Sodium is dissolved in methanol to generate sodium methoxide. S25-42 isadded and the mixture stirred at 62° C. for 16 hr. The mixture is cooledand acetic acid added with cooling to neutralize the excess sodiummethoxide. The mixture is diluted with water and extracted with ethylacetate. The extract is dried and evaporated to give S25-43.

Example 46Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-methyl-5-exo-(t-butoxycarbonyl)aminodicarboxylate (S25-44)

Compound S25-43 is reacted with benzyl bromide and cesium carbonate intetrahydrofuran at room temperature to give benzyl ester S25-44, whichis isolated by acid work-up of the crude reaction mixture.

Example 47Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-carboxy-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S25-45)

Compound S25-44 is dissolved in methanol and cooled to 0° C. under N₂.2M NaOH (2 equivalents) is added dropwise, the mixture allowed to cometo ambient temperature and is stirred for 5 h. The solution is dilutedwith water, acidified with 2M HCl and extracted with ethyl acetate. Theextract is washed with water, saturated aqueous NaCl, dried andevaporated to give S25-45.

Example 48Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-(trimethylsilylethoxycarbonyl)amino-5-exo-(t-butoxycarbonyl)aminocarboxylate (S25-46)

To a solution of S25-45 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours and thencooled to room temperature. Trimethylsilylethanol is added and thesolution refluxed for 48 hours. The solution is partitioned betweenethyl acetate and 1M sodium bicarbonate. The organic layer is washedwith 1M sodium bicarbonate and dried over sodium sulfate. The solvent isevaporated under reduced pressure to give crude Curtius product S25-46.

Example 49endo-Bicyclo[2.2.1]hept-5-ene-2-(4-methoxy)benzylcarboxylate-3-carboxylicacid (S26-48)

Compound S23-19 and quinidine are suspended in equal amounts of tolueneand carbon tetrachloride and cooled to −55° C. p-Methoxybenzyl alcoholis added over 15 minutes and the solution stirred at −55° C. for 96hours. After removal of the solvents, the residue is partitioned betweenethyl acetate and 2 M hydrochloric acid. The organic layer is washedwith water and saturated aqueous sodium chloride. Drying over magnesiumsulfate and removal of the solvent gives S26-48.

Example 50endo-Bicyclo[2.2.1]hept-5-ene-2-(4-methoxy)benzyl-3-(trimethylsilylethoxy-carbonyl)aminocarboxylate (S26-49)

To a solution of S26-48 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours, cooledto room temperature, trimethylsilylethanol is added, and the solution isrefluxed for an additional 48 hours. The benzene solution is partitionedbetween ethyl acetate and 1 M sodium bicarbonate. The organic layers arecombined, washed with 1 M sodium bicarbonate, and dried with sodiumsulfate. The solvent is evaporated under reduced pressure to give crudeCurtius product S26-49.

Example 51Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-methyl-6-exo-methyltricarboxylate (S26-50)

S26-49, copper(II) chloride, 10% Pd/C, and dry methanol are added to aflask with vigorous stirring. After degassing the suspension, the flaskis charged with carbon monoxide to a pressure just above 1 atm. Thepressure of carbon monoxide is maintained over 72 hours. The solids arefiltered off, and the crude reaction mixture worked up in the usual wayto afford S26-50.

Example 52Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-6-exo-dicarboxylicanhydride (S26-51)

S26-50, formic acid, and a catalytic amount of p-toluenesulfonic acid isheated at 90° C. overnight. Acetic anhydride is added to the reactionmixture, and it is refluxed for an additional 6 hours. Removal of thesolvents and washing with ether affords S26-51.

Example 53Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-carboxy-6-exo-methyldicarboxylate (S26-52)

To a solution of S26-51 in equal amounts of toluene and carbontetrachloride is added quinine. The suspension is cooled to −65° C. andstirred for 1 hour. Three equivalents of methanol are added slowly over30 minutes. The suspension is stirred at −65° C. for 4 days followed byremoval of the solvents. The resulting white solid is partitionedbetween ethyl acetate and 2 M HCl, with S26-52 worked up from theorganic layer.

Example 54Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-(trimethylsilylethoxycarbonyl)amino-6-exo-methyldicarboxylate (S26-53)

To a solution of S26-52 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours thencooled to room temperature. 2-Trimethylsilylethanol is added, and thesolution is refluxed for an additional 48 hours. The benzene solution ispartitioned between ethyl acetate and 1 M sodium bicarbonate. Theorganic layers are combined, washed with 1 M sodium bicarbonate, anddried with sodium sulfate. The solvent is evaporated under reducedpressure to give S26-53.

Example 55Bicyclo[2.2.1]heptane-2-exo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-(trimethylsilylethoxycarbonyl)amino-6-endo-methyldicarboxylate (S26-54)

To a solution of S26-53 in tetrahydrofuran is carefully added potassiumtert-butoxide. The basic solution is refluxed for 24 hours followed byaddition of acetic acid. Standard extraction methods give the doubleepimerized product S26-54.

Example 56

Preparation of hexamer:

To 0.300 g (1R,2R)-(−)-trans-1,2-diaminocyclohexane (2.63 mmol) in 5 mLCH₂Cl₂ at 0° C. was added 0.600 g of 2,6-diformyl-4-bromophenol (2.62mmol) in 5 mL of CH₂Cl₂. The yellow solution was allowed to warm to roomtemperature and stirred for 48 hours. The reaction solution wasdecanted, and added to 150 mL of methanol. After standing for 30minutes, the yellow precipitate was collected, washed with methanol, andair-dried (0.580 g; 72% yield).

¹H NMR (400 MHz, CDCl₃) δ 14.31 (s, 3H, OH), 8.58 (s, 3H, CH═N), 8.19(s, 3H, CH═N), 7.88 (d, 3H, J=2.0 Hz, ArH), 7.27 (d, 3H, J=2.0 Hz, ArH),3.30-3.42 (m, 6H, CH₂—CH—N), 1.41-1.90 (m, 24H, aliphatic).

MS (FAB): Calcd for C₄₂H₄₆N₆O₃Br₃ 923.115; found 923.3 [M+H]⁺.

Example 57

Preparation of hexamer:

To 0.300 g (1R,2R)-(−)-trans-1,2-diaminocyclohexane (2.63 mmol) in 6 mLCH₂Cl₂ at 0° C. was added 0.826 g of2,6-diformyl-4-(1-dodec-1-yne)phenol (2.63 mmol) in 6 mL of CH₂Cl₂. Theorange solution was stirred at 0° C. for 1 hour and then allowed to warmto room temperature after which stirring was continued for 16 hours. Thereaction solution was decanted and added to 150 mL of methanol. A stickyyellow solid was obtained after decanting the methanol solution.Chromatographic cleanup of the residue gave a yellow powder.

¹H NMR (400 MHz, CDCl₃) δ 14.32 (s, 3H, OH), 8.62 (s, 3H, CH═N), 8.18(s, 3H, CH═N), 7.84 (d, 3H, J=2.0 Hz, ArH), 7.20 (d, 3H, J=2.0 Hz, ArH),3.30-3.42 (m, 6H, CH₂—CH—N), 2.25 (t, 6H, J=7.2 Hz, propargylic),1.20-1.83 (m, 72H, aliphatic), 0.85 (t, 9H, J=7.0 Hz, CH₃).

¹³C NMR (400 MHz, CDCl₃) δ 163.4, 161.8, 155.7, 136.9, 132.7, 123.9,119.0, 113.9, 88.7, 79.7, 75.5, 73.2, 33.6, 33.3, 32.2, 29.8, 29.7,29.6, 29.4, 29.2, 29.1, 24.6, 24.5, 22.9, 19.6, 14.4.

MS (FAB): Calcd for C₇₈H₁₀₉N₆O₃ 1177.856; found: 1177.8 [M+H]⁺.

Example 58

Preparation of hexamer:

To 0.240 g of 2,6-diformyl-4-(1-dodecene)phenol (0.76 mmol) in 10 mL ofbenzene was added a 10 mL benzene solution of(1R,2R)-(−)-trans-1,2-diaminocyclohexane (0.087 g, 0.76 mmol). Thesolution was stirred at room temperature for 48 hours shielded from thelight. The orange solution was taken to dryness and chromatographed(silica, 50/50 acetone/Et₂O) to give a yellow sticky solid (33% yield).

¹H NMR (400 MHz, CDCl₃) δ 14.12 (s, 3H, OH), 8.62 (s, 3H, CH═N), 8.40(s, 3H, CH═N), 7.82 (d, 3H, J=2.0 Hz, ArH), 7.28 (d, 3H, J=2.0 Hz, ArH),6.22 (d, 3H, vinyl), 6.05 (d, 3H, vinyl), 3.30-3.42 (m, 6H, CH₂—CH—N),1.04-1.98 (m, 87H, aliphatic).

MS (FAB): Calcd for C₇₈H₁₁₅N₆O₃ 1183.90; found: 1184.6 [M+H]⁺.

Example 59

Preparation of tetramer:

Preparation of hexamer:

Triethylamine (0.50 mL, 3.59 mmol) and(1R,2R)-(−)-trans-1,2-diaminocyclohexane (0.190 g, 1.66 mmol) werecombined in 150 mL EtOAc and purged with N₂ for 5 minutes. To thissolution was added 0.331 g isophthalolyl chloride (1.66 mmol) dissolvedin 100 mL EtOAc dropwise over six hours. The solution was filtered andthe filtrate taken to dryness. TLC (5% methanol/CH₂Cl₂) shows theproduct mixture to be primarily composed of two macrocycliccompositions. Chromatographic separation (silica, 5% methanol/CH₂Cl₂)gave the above tetramer (0.020 g, 5% yield) and hexamer (about 10%).

Tetramer:

¹H NMR (400 MHz, CDCl₃) δ 7.82 (s, 1H), 7.60 (br s, 2H), 7.45 (br s,2H), 7.18 (br s, 1H), 3.90 (br s, 2H), 2.22 (d, 2H), 1.85 (m, 4H), 1.41(m, 4H).

MS (ESI): Calcd for C₂₈H₃₃N₄O₄ 489.25; found 489.4 [M+H]⁺.

Hexamer:

MS (ESI): Calcd for C₄₂H₄₉N₆O₆ 733.37; found 733.5 [M+H]⁺.

Example 60

Preparation of macrocyclic modules from benzene and cyclohexane cyclicsynthons:

To a 5 mL dichloromethane solution of 4-dodecyl-2,6-diformyl anisole (24mg; 0.072 mmol) was added a 5 mL dichloromethane solution of(1R,2R)-(−)-trans-1,2-diaminocyclohexane (8.5 mg; 0.074 mmol). Thissolution was stirred at room temperature for 16 hours and then added tothe top of a short silica column. Elution with diethyl ether and thenremoval of solvent led to the isolation of 22 mg of an off-white solid.Positive ion electrospray mass spectrometry demonstrated the presence ofthe tetramer (m/z 822, MH⁺), hexamer (m/z 1232, MH⁺), and the octamer(m/z 1643, MH⁺) in the off-white solid. Calculated molecular weightswere as follows: tetramer+H (C₅₄H₈₅N₄O₂, 821.67); hexamer+H(C₈₁H₁₂₇N₆O₃, 1232.00); octamer+H (C₁₀₈H₁₆₉N₈O₄, 1643.33).

Example 61

Templated Imine Octamer. To a 3 neck 100 mL round bottomed flask withstirbar, fitted with condenser and addition funnel under argon,amphiphilic dialdehyde phenol 1 (500 mg, 1.16 mmol) was added. Next,Mg(NO₃)₂.6H₂O (148 mg, 0.58 mmol) 2 and Mg(OAc)₂.4H₂O (124 mg, 0.58mmol) were successively added. The flask was put under vacuo andbackfilled with argon 3×. Anhydrous methanol was transferred to theflask via syringe under argon and the resulting suspension stirred. Themixture was then refluxed for 10 min affording a homogeneous solution.The reaction was allowed to cool to room temperature under positiveargon pressure. (1R,2R)-(−)-trans-1,2-diaminocyclohexane 4 was added tothe addition funnel followed by cannula transfer of anhydrous MeOH (11.6mL) under argon. The diamine/MeOH solution was added to the stirredhomogeneous metal template/dialdehyde solution drop wise over a periodof 1 h affording an orange oil. The addition funnel was replaced with aglass stopper and the mixture was refluxed for 3 days. The solvent wasremoved in vacuo affording a yellow crystalline solid that was usedwithout further purification.

Amine Octamer. To a 50 mL schlenk flask with stirbar under argon ImineOctamer (314 mg, 0.14 mmol) was added. Next anhydrous THF (15 mL) andMeOH (6.4 mL) were added via syringe under argon and the suspensionstirred at room temperature. To the homogeneous solution, NaBH₄ (136 mg,3.6 mmol) was added in portions and the mixture stirred at roomtemperature for 12 h. The solution was filtered, followed by addition of19.9 mL H₂O. The pH was adjusted to ca. 2 by addition of 4 M HCl, then6.8 mL of an ethylenediamine tetraacetic acid disodium salt dihydrate(0.13 M in H₂O) was added and the mixture stirred for 5 min. To thesolution, 2.0% ammonium hydroxide was added and stirring continued foran additional 5 min. The solution was extracted with ethyl acetate(3×100 mL) the organic layer separated, dried over Na₂SO₄ and thesolvent removed via rotoevaporation affording a pale yellow solid.Recrystallization from chloroform and hexanes afforded the AmineOctamer. Molecular weight was confirmed by ESIMS M+H=experimental=2058.7m/z, calcd=2058.7 m/z.

Example 62

Hexamer 1j. The two substrates, (−)-R,R-1,2-trans-diaminocyclohexane(0.462 mmol, 0.053 g) and 2,6-diformyl-4-hexadecyl benzylphenolcarboxylate (0.462 mmol, 0.200 g) were added to a 10 mL vial containinga magnetic stirbar followed by the addition of 2 mL of CH₂Cl₂. Theyellow solution was stirred at room temperature. After 24 h the reactionsolution was plugged through silica gel with diethyl ether, and thesolvent removed via roto-evaporation. (232 mg; 98% yield). ¹H NMR (400MHz, CDCl₃): δ 14.11 (s, 3H, OH), 8.67 (s, 3H, CH═N), 8.23 (s, 3H,CH═N), 7.70 (s, 3H, ArH), 7.11 (s, 3H, ArH), 4.05-3.90 (t, 6H, ³J=6.6Hz, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.44 (s, 6H, CH₂C(O)OCH₂(CH₂)₁₄CH₃),3.30-3.42 (m, 6H, CH₂—CH—N), 1.21-1.90 (m, 108H, aliphatic) 0.92-0.86(t, 9H, ³J=6.6 Hz. ESIMS (+) Calcd for C₉₆H₁₅₁N₆O₉: 1533; Found: 1534[M+H]⁺.

Hexamer 1jh. To a 100 mL pear-shaped flask with magnetic stirbar underargon, Hexamer 1j (0.387 mmol, 0.594 g) was added and dissolved inTHF:MeOH (7:3, 28:12 mL, respectively). Next, NaBH₄ (2.32 mmol, 0.088 g)was added slowly in portions at room temperature for 6.5 h. The solventwas removed by roto-evaporation, the residue dissolved in 125 mL ethylacetate and washed 3×50 mL of H₂O. The organic layer was separated,dried over Na₂SO₄ and the solvent removed by roto-evaporation. Theresulting residue was recrystallized from CH₂Cl₂ and MeOH affording awhite solid (0.440 g; 74% yield). ¹H NMR (400 MHz, CDCl₃): δ 6.86 (s,6H, ArH), 4.10-4.00 (t, 6H, ³J=6.6 Hz, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.87-3.69(dd, 6H, ³J=13.7 Hz, ³J (CNH)=42.4 Hz CH₂—CH—N), 3.43 (s, 6H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 2.40-2.28 (m, 6H, aliphatic), 2.15-1.95 (m, 6H,aliphatic), 1.75-1.60 (m, 6H, aliphatic), 1.60-1.55 (m, 6H, aliphatic)1.37-1.05 (m, 84H, aliphatic) 0.92-0.86 (t, 9H, ³J=6.8 Hz. ESIMS (+)Calcd for C₉₆H₁₆₃N₆O₉: 1544; Found: 1545 [M+H]⁺.

Example 63

Hexamer 1A-Me. A solution of2-hydroxy-5-methyl-1,3-benzenedicarboxaldehye (53 mg, 0.32 mmol) indichloromethane (0.6 mL) was added to a solution of(1R,2R)-(−)-1,2-diaminocyclohexane (37 mg, 0.32 mmol) in dichloromethane(0.5 mL). The mixture was stirred at ambient temperature for 16 h, addeddropwise to methanol (75 mL) and chilled (4° C.) for 4 h. Theprecipitate was collected to afford 71 mg (92%) of hexamer 1A-Me. ¹H NMR(CDCl₃): δ 13.88 (s, 3H, OH), 8.66 (s, 3H, ArCH═N), 8.19 (s, 3H,ArCH═N), 7.52 (d, 3H, J=2 Hz, Ar H), 6.86 (d, 3H, J=2 Hz, Ar H), 3.35(m, 6H, cyclohexane 1,2-H's), 2.03 (3, 9H, Me), 1.6-1.9 (m, 18H,cyclohexane 3,6-H₂ and 4_(eq), 5_(eq)-H's), 1.45 (m, 6H, cyclohexane4_(ax), 5_(ax)-H's); ¹³C NMR δ 63.67, 159.55, 156.38, 134.42, 129.75,127.13, 119.00, 75.68, 73.62, 33.68, 33.41, 24.65, 24.57, 20.22; ESI(+)MS m/e (%) 727 M+H (100); IR 1634 cm⁻¹.

Example 64

32.7 mg Hexamer 1jh (recrystallized times) was added to 30 mL dry THF.100 μL triethylamine and 100 μL acryloyl chloride (freshly distilled)were added subsequently to the THF mixture using Schlenk technique.Solution was stirred for 18 hrs in an acetone/dry ice bath. Afterremoval of solvent a white precipitate remained. The precipitate wasredissolved in CH₂Cl₂ and filtered through a fritted funnel. CH₂Cl₂solution was added to the separatory funnel and washed one time withwater followed by two brine (NaCl) washes. The CH₂Cl₂ solution was driedover MgSO₄ and then filtered to remove MgSO₄. A yellow precipitateremained after solvent removal. ¹H NMR (CDCl₃): δ −0.867-0.990 (3H),1.259 (21.8H), 1.39 (1.86H), 1.64 (12.7H), 2.8 (1.25H), 3.46-3.62(2.47H), 3.71 (0.89H), 3.99 (2.46H), 5.06 (0.71H), 5.31 (3.80H), 5.71(1.43H), 5.90 (0.78H), 6.2-6.4 (2.49H), 6.59 (0.80H), 6.78 (0.47H), 6.98(0.28H). FTIR-ATR: 3340, 2926 (—CH₂—), 2854 (—CH₂—), 1738 (EsterCarbonyl), 1649 and 1613 (Acrylate), 983 (═CH), 959 sh (═CH₂). ESI-MS:1978.5 (Hex1JhAC+8-AC), 1948.8 (Hex1JhAC+7-AC+Na⁺), 1923.3(Hex1JhAC+7-AC), 1867.6 (Hex1JhAC+6-AC), 1842.6, 1759.7 (Hex1JhAC+4-AC).

We claim:
 1. A bridged macrocyclic module compound of the formula:

wherein the compound further comprises a bridge moiety A having two ormore termini, wherein at least two of said two or more termini arecoupled to the compound; wherein each Q is a synthon independentlyselected from the group consisting of: benzene, cyclohexadiene,cyclopentadiene, naphthalene, anthracene, phenylene, phenanthracene,pyrene, triphenylene, phenanthrene, pyridine, pyrimidine, pyridazine,biphenyl, bipyridyl, cyclohexane, cyclohexene, decalin, piperidine,pyrrolidine, morpholine, piperazine, pyrazolidine, quinuclidine,tetrahydropyran, dioxane, tetrahydrothiophene, tetrahydrofuran, pyrrole,cyclopentane, cyclopentene, triptycene, adamantane,bicyclo[2.2.1]heptane, bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane,bicyclo[2.2.2]octene, bicyclo[3.3.0]octane, bicyclo[3.3.0]octene,bicyclo[3.3.1]nonane, bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane,bicyclo[3.2.2]nonene, bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, spiro[4.4]nonane, —OCH₂CH₂—,—(CH₂)_(n)C≡C(CH₂)_(n)—, —(CH₂)_(n)CH═CH(CH₂)_(n)—,

—(CH₂)_(n)—, —C(O)O(CH₂)_(n)—, —(CH₂)_(n)C(O)NR—; —S_(m)—,—(CH₂)_(n)SiMe₂(CH₂)_(n)—, —(CH₂)_(n)NR(CH₂)_(n)—, and—(CH₂)_(n)CH(OH)—; wherein each synthon Q may optionally be substitutedwith one or more functional groups for coupling the synthon to at leasta second bridged macrocyclic module or to a substrate; wherein eachsynthon may optionally be substituted with one or more lipophilic and/orhydrophilic groups; wherein each L is a linkage moiety independentlyselected from the group consisting of a direct bond, —NRC(O)—, —OC(O)—,—O—, —S—S—, —S—, —NR—, —(CRR)_(p)—, —CH₂NH—, —CH═N—, —C(O)S—, —C(O)O—,—C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—,—NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—N═CH(CH₂)_(h)CH═N—, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—,—CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

wherein the linkage is independently configured in either of twopossible configurations, forward and reverse, with respect to thesynthons it couples together; wherein the bridge moiety A is selectedfrom the group consisting of

—O—(CH₂)_(m)—O—; —{NH—CHR—(CO)}_(m)—O—; —O—(CF₂)_(m)—O—; —(S)_(m)—;—O(CH₂CH₂O)_(m)—; —(OCH(CH₃)CH₂)_(m)O—;

wherein the at least two termini of the bridge moiety may be conjugatedto the compound through a linkage moiety L, wherein L is as definedabove; wherein R is independently selected from the group consisting ofhydrogen and alkyl; wherein Ph is phenyl; wherein X is selected from thegroup consisting of F, Cl, Br, and I; wherein X′ is independently H or afunctional group for linking to at least a second bridged macrocyclicmoiety or a substrate; wherein each R² is independently selected from abond for linking to a synthon or a functional group selected from thegroup consisting of hydrogen, an activated acid, —OH, —C(O)OH, —C(O)H,—C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa, —SH,—C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂,—CH═CHR, —CH═CRR, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

—P(O)(OH)(OX), or —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein n′ is from 4 to 50;wherein n is 1-22; wherein m is 2-14; wherein p is 1-6; wherein h is1-4; wherein r is 1-50; and wherein s is 1-4.
 2. The compound of claim1, wherein the two or more termini of said bridge moiety are coupled tosynthons.
 3. The compound of claim 1, wherein the two or more termini ofsaid bridge moiety are coupled to L moieties, with the proviso that saidL moieties to which the termini are coupled are not direct bonds.
 4. Thecompound of claim 1, wherein n′ is from 4 to
 24. 5. The compound ofclaim 1, wherein n′ is from 6 to
 12. 6. The compound of claim 1, havingthe formula:

wherein each Q¹, Q², Q³, and L are independently selected.
 7. Thecompound of claim 1, having the formula:

wherein each Q¹, Q², Q³, and L are independently selected.
 8. Thecompound of claim 1, having the formula:

wherein each Q¹, Q², Q³, and L are independently selected.
 9. Thecompound of claims 6-8, wherein each Q¹ is the same synthon.
 10. Thecompound of claims 6-8, wherein each Q² is the same synthon.
 11. Thecompound of claims 6-8, wherein each Q³ is the same synthon.
 12. Thecompound of claims 6-8, wherein each Q¹, Q², and Q³ is independentlyselected from the group consisting of

wherein each X′ is independently H or a functional group for couplingthe synthon to at least a second bridged macrocyclic module or to asubstrate; wherein each J is an independently selected functional groupfor coupling the synthon to an adjacent synthon within said bridgedmacrocyclic module, and wherein each X₁ is an independently selectedfunctional group for coupling the synthon to the bridge moiety.
 13. Thecompound of claim 12, wherein each Q¹, Q², and Q³ is independentlyselected from the group consisting of


14. The compound of claim 12, wherein each Q¹, Q², and Q³ isindependently selected from the group consisting of


15. The compound of claims 6-8, wherein each L between the synthons arethe same.
 16. The compound of claims 6-8, wherein each L between thebridge moiety and the synthons are the same.
 17. The compound of claim1, wherein the synthons are cyclic synthons.
 18. The compound of claim1, wherein the synthons are acyclic synthons.
 19. The compound of claim1, wherein each L is a direct bond.
 20. The compound of claim 1, whereineach L is a linkage independently selected from the group consisting of—NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR)_(p)—, —CH₂NH—, —C(O)S—,—C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—,—NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—NH(CH₂)_(h)CH═N—, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—,—CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,


21. The compound of claim 1, wherein the bridge moiety further comprisesa surface attachment group.
 22. The compound of claim 1, wherein thebridge moiety further comprises a lipophilic group.
 23. The compound ofclaim 1, wherein the bridge moiety comprises a functional group forcoupling the compound to at least a second bridged macrocyclic modulecompound.
 24. The compound of claim 1, wherein the bridge moietycomprises a polymerization center.
 25. The compound of claim 1, whereinthe bridged macrocyclic module compound is selected from the groupconsisting of:

wherein R^(o) is H, alkyl, or a lipophilic group; wherein R′ is anatural α-amino acid side chain; wherein R¹ is CH₂CO₂(CH₂)₁₅CH₃; andwherein the structure

is benzene or cyclohexane.
 26. A nanofilm comprising a plurality ofbridged macrocyclic modules of claim
 1. 27. The nanofilm of claim 26,wherein the thickness of the nanofilm composition is less than about 30nanometers.
 28. The nanofilm of claim 26, wherein the thickness of thenanofilm composition is less than about 6 nanometers.
 29. The nanofilmof claim 26, wherein the nanofilm is impermeable to viruses and largerspecies.
 30. The nanofilm of claim 26, wherein the nanofilm isimpermeable to immunoglobulin G and larger species.
 31. The nanofilm ofclaim 26, wherein the nanofilm is impermeable to albumin and largerspecies.
 32. The nanofilm of claim 26, wherein the nanofilm isimpermeable to β2-Microglobulin and larger species.
 33. The nanofilm ofclaim 26, wherein the nanofilm is permeable only to water and smallerspecies.
 34. The nanofilm of claim 26, wherein the nanofilm has amolecular weight cut-off of 13 kDa.
 35. The nanofilm of claim 26,wherein the nanofilm has a molecular weight cut-off of 190 Da.
 36. Thenanofilm of claim 26, wherein the nanofilm has a molecular weightcut-off of 100 Da.
 37. The nanofilm of claim 26, wherein the nanofilmhas a molecular weight cut-off of 45 Da.
 38. The nanofilm of claim 26,wherein the nanofilm has a molecular weight cut-off of 20 Da.
 39. Thenanofilm of claim 26, wherein the nanofilm has high permeability forwater molecules and Na+, K+, and Cs+ in water.
 40. The nanofilm of claim26, wherein the nanofilm has low permeability for glucose and urea. 41.The nanofilm of claim 26, wherein the nanofilm has high permeability forwater molecules and Cl− in water.
 42. The nanofilm of claim 26, whereinthe nanofilm has high permeability for water molecules and K+ in water,and low permeability for Na+ in water.
 43. The nanofilm of claim 26,wherein the nanofilm has high permeability for water molecules and Na+in water, and low permeability for K+ in water.
 44. The nanofilm ofclaim 26, wherein the nanofilm has low permeability for urea,creatinine, Li+, Ca2+, and Mg2+ in water.
 45. The nanofilm of claim 26,wherein the nanofilm has high permeability for Na+, K+, hydrogenphosphate, and dihydrogen phosphate in water.
 46. The nanofilm of claim26, wherein the nanofilm has high permeability for Na+, K+, and glucosein water.
 47. The nanofilm of claim 26, wherein the nanofilm has lowpermeability for myoglobin, ovalbumin, and albumin in water.
 48. Thenanofilm of claim 26, wherein the nanofilm has high permeability fororganic compounds and low permeability for water.
 49. The nanofilm ofclaim 26, wherein the nanofilm has low permeability for organiccompounds and high permeability for water.
 50. The nanofilm of claim 26,wherein the nanofilm has low permeability for water molecules and highpermeability for helium and hydrogen gases.
 51. A nanofilm compositioncomprising at least two layers of the nanofilm of claim
 26. 52. Thenanofilm of claim 51, wherein the nanofilm composition comprises atleast one spacing layer between any two of the nanofilm layers.
 53. Thenanofilm of claim 52, wherein the spacing layer comprises a layer of apolymer, a gel, or inorganic particles.
 54. The nanofilm of claim 26,wherein the nanofilm is deposited on a substrate.
 55. The nanofilm ofclaim 54, wherein the substrate is porous.
 56. The nanofilm of claim 54,wherein the nanofilm is coupled to the substrate throughbiotin-strepavidin mediated interaction.
 57. A method of filtration,comprising using a nanofilm of claim 26 to separate components fromfluid.
 58. A method of making a bridged macrocyclic module compound ofclaim 1, comprising: (a) providing a bridged program director compoundof the structure

wherein Q¹ is a synthon; wherein A is a bridge moiety; wherein A isconjugated to Q¹ through a linkage moiety L; wherein each J¹ is afunctional group for coupling an adjacent synthon; and wherein n′ is4-25; and (b) reacting a synthon or a synthon multimer with said bridgedprogram director compound to form a bridged macrocyclic module compound.59. A method of making a bridged macrocyclic module compound of claim 1,comprising: (a) providing a macrocyclic moiety compound, wherein themacrocyclic moiety compound contains from 4 to 50 synthons; and (b)reacting a bridge moiety comprising at least two termini with saidmacrocyclic moiety compound to form a bridged macrocyclic modulecompound.